Compositions comprising bacterial strains

ABSTRACT

The invention provides compositions comprising bacterial strains for treating and preventing a neurodegenerative disorder.

CROSS-REFERENCE

This application is a continuation of International Application No.PCT/EP2018/065808, filed Jun. 14, 2018, which claims the benefit ofGreat Britain Application No. 1709466.5, filed Jun. 14, 2017, and GreatBritain Application No. 1709533.2, filed Jun. 15, 2017 all of which arehereby incorporated by reference in their entirety.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ANSI format and is hereby incorporated byreference in its entirety. Said ANSI copy, created on Jan. 20, 2020, isnamed 56708-728_301_SL.txt and is 4,680,747 bytes in size.

TECHNICAL FIELD

This invention is in the field of compositions comprising bacterialstrains isolated from the mammalian digestive tract and the use of suchcompositions in the treatment of disease.

BACKGROUND TO THE INVENTION

The human intestine is thought to be sterile in utero, but it is exposedto a large variety of maternal and environmental microbes immediatelyafter birth. Thereafter, a dynamic period of microbial colonization andsuccession occurs, which is influenced by factors such as delivery mode,environment, diet and host genotype, all of which impact upon thecomposition of the gut microbiota, particularly during early life.Subsequently, the microbiota stabilizes and becomes adult-like [1]. Thehuman gut microbiota contains more than 500-1000 different phylotypesbelonging essentially to two major bacterial divisions, theBacteroidetes and the Firmicutes [2]. The successful symbioticrelationships arising from bacterial colonization of the human gut haveyielded a wide variety of metabolic, structural, protective and otherbeneficial functions. The enhanced metabolic activities of the colonizedgut ensure that otherwise indigestible dietary components are degradedwith release of by-products providing an important nutrient source forthe host. Similarly, the immunological importance of the gut microbiotais well-recognized and is exemplified in germfree animals which have animpaired immune system that is functionally reconstituted following theintroduction of commensal bacteria [3-5].

Dramatic changes in microbiota composition have been documented ingastrointestinal disorders such as inflammatory bowel disease (IBD). Forexample, the levels of Clostridium cluster XIVa bacteria are reduced inIBD patients whilst numbers of E. coli are increased, suggesting a shiftin the balance of symbionts and pathobionts within the gut [6-9].

In recognition of the potential positive effect that certain bacterialstrains may have on the animal gut, various strains have been proposedfor use in the treatment of various diseases (see, for example,[10-13]). Also, certain strains, including mostly Lactobacillus andBifidobacterium strains, have been proposed for use in treating variousinflammatory and autoimmune diseases that are not directly linked to theintestines (see [14] and [15] for reviews). The ability of Roseburiahominis to regulate the immune system has been suggested in [16].However, the relationship between different diseases and differentbacterial strains, and the precise effects of particular bacterialstrains on the gut and at a systemic level and on any particular typesof diseases are poorly characterised, particularly for neurodegenerativedisorders.

Recently, there has been increased interest in the art regardingalterations in the gut microbiome that may play a pathophysiologicalrole in human brain diseases [17]. Preclinical and clinical evidence arestrongly suggesting a link between brain development and microbiota[18]. A growing body of preclinical literature has demonstratedbidirectional signalling between the brain and the gut microbiome,involving multiple neurocrine and endocrine signalling systems. Indeed,increased levels of Clostridium species in the microbiome have beenlinked to brain disorders [19], and an imbalance of the Bacteroidetesand Finnicutes phyla has also been implicated in brain developmentdisorders [20]. Suggestions that altered levels of gut commensals,including those of Bifidobacterium, Lactobacillus, Sutterella,Prevotella and Ruminococcus genera and of the Alcaligenaceae family areinvolved in immune-mediated central nervous system (CNS) disorders, arequestioned by studies suggesting a lack of alteration in the microbiotabetween patients and healthy subjects [10]. Roseburia hominis has beenproposed for treating a variety of disorders including asthma,rheumatoid arthritis and multiple sclerosis [21]

Like asthma and rheumatoid arthritis, multiple sclerosis is primarilymediated by the immune system. The immune system attacks myelinatedaxons in the central nervous system, destroying the myelin calledplaques or lesions. Demyelination occurs in particular in the opticnerves, subpial spinal cord, brainstem, cerebellum, and juxtacorticaland periventricular white matter regions.

As such, multiple sclerosis has a different pathology to otherneurodegenerative diseases, such as Parkinson's disease, Alzheimer'sdisease or dementia. For example, multiple sclerosis is commonlydiagnosed in patients in their 20s and 30s, while many otherneurodegenerative diseases, such as Parkinson's disease, Alzheimer's anddementia, are diagnosed predominantly in patients aged over 65 yearsold.

Parkinson's disease, like many neurodegenerative diseases, is primarilymediated by the accumulation of misfolded protein. Parkinson's diseaseis a synucleinopathology that involves the accumulation of α-synuclein,which aggregate as insoluble fibrils in Lewy bodies within the cytoplasmof the neuronal body. The accumulation of α-synuclein is toxic andimpairs the functions of mitochondria, lysosomes, and endoplasmicreticulum, and interferes with microtubule transport.

Nonsteroidal anti-inflammatory drugs (NSAIDs) such as ibuprofen, havebeen tested for their efficacy in treating a variety of neurologicaldiseases, but the clinical impact of NSAIDs on neurodegenerativediseases like Parkinson's disease remains unclear. While some studiesshowed that chronic NSAID use is protective against Parkinson's disease,other studies could not confirm the existence of a significantrelationship. A recent meta-analysis indicated that the use ofnon-aspirin NSAID, particularly ibuprofen, reduces the risk of PD by 15%while the use of aspirin did not show any effect [22].

This indicates that, at present, the practical effect of the linkbetween the microbiome and human brain diseases is poorly characterised.Accordingly, more direct analytical studies are required to identify thetherapeutic impact of altering the microbiome on neurodegenerativedisorders.

There is a requirement in the art for new methods of treatingneurodegenerative disorders. There is also a requirement for thepotential effects of gut bacteria to be characterised so that newtherapies using gut bacteria can be developed.

SUMMARY OF THE INVENTION

The inventors have developed new therapies for treating and preventingneurodegenerative disorders. The inventors have identified thatbacterial strains from the genus Roseburia may be effective for treatingneurodegenerative diseases. As described in the examples, administrationof compositions comprising Roseburia hominis can protect againstreactive oxygen species and prevent inflammation, thus acting as aneuroprotectant. The inventors have also identified that treatment withRoseburia hominis can reduce the activation of proinflammatorymolecules, such as NFκB and IL-6, by LPS and mutant α-synuclein A53T.The inventors have identified that treatment with Roseburia hominis canreduce histone deacetylation activity and lipid peroxidation in vitro,which can help to reduce cell death and apoptosis. The inventors havealso identified that Roseburia hominis can produce indole that canattenuate inflammation and oxidative stress. Furthermore, the inventorshave demonstrated that treatment with Roseburia hominis can increasekynurenine levels.

In a first embodiment, the invention provides a composition comprising abacterial strain of the genus Roseburia, for use in a method of treatingor preventing a neurodegenerative disorder.

In particular embodiments, the invention provides a compositioncomprising a bacterial strain of the genus Roseburia, for use in amethod of treating or preventing a disease or condition selected fromthe group consisting of: Parkinson's disease, including progressivesupranuclear palsy, progressive supranuclear palsy,Steele-Richardson-Olszewski syndrome, normal pressure hydrocephalus,vascular or arteriosclerotic parkinsonism and drug-induced parkinsonism;Alzheimer's disease, including Benson's syndrome; multiple sclerosis;Huntington's disease; amyotrophic lateral sclerosis; Lou Gehrig'sdisease; motor neurone disease; prion disease; spinocerebellar ataxia;spinal muscular atrophy; dementia, including Lewy body, vascular andfrontotemporal dementia; primary progressive aphasia; mild cognitiveimpairment; HIV-related cognitive impairment and corticobasaldegeneration.

In preferred embodiments, the invention provides a compositioncomprising a bacterial strain of the genus Roseburia, for use in amethod of treating or preventing Parkinson's disease, such asenvironmental, familial or Parkinson's associated with generalinflammatory status. The inventors have identified that treatment withRoseburia strains can reduce the activation of proinflammatorymolecules, such as NFκB and IL-6, by LPS and mutant α-synuclein A53T inin vitro models of environmental and familial Parkinson's. In preferredembodiments, the invention provides a composition comprising a bacterialstrain of the species Roseburia hominis, for use in the treatment ofParkinson's disease. Compositions using Roseburia hominis may beparticularly effective for treating Parkinson's.

In preferred embodiments of the invention, the bacterial strain in thecomposition is of Roseburia hominis. Closely related strains may also beused, such as bacterial strains that have a 16SrRNA sequence that is atleast 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% identical to the 16S rRNAsequence of a bacterial strain of Roseburia hominis. Preferably, thebacterial strain has a 16S rRNA sequence that is at least 95%, 96%, 97%,98%, 99%, 99.5% or 99.9% identical to SEQ ID NO:1, 2 or 3. Preferably,the sequence identity is to SEQ ID NO:3. Preferably, the bacterialstrain for use in the invention has the 16S rRNA sequence represented bySEQ ID NO:3.

In certain embodiments, the composition of the invention is for oraladministration. Oral administration of the strains of the invention canbe effective for neurodegenerative disorders. Also, oral administrationis convenient for patients and practitioners and allows delivery toand/or partial or total colonisation of the intestine.

In certain embodiments, the composition of the invention comprises oneor more pharmaceutically acceptable excipients or carriers.

In certain embodiments, the composition of the invention comprises abacterial strain that has been lyophilised. Lyophilisation is aneffective and convenient technique for preparing stable compositionsthat allow delivery of bacteria.

In certain embodiments, the invention provides a food product comprisingthe composition as described above.

In certain embodiments, the invention provides a vaccine compositioncomprising the composition as described above.

Additionally, the invention provides a method of treating or preventingneurodegenerative disorders, comprising administering a compositioncomprising a bacterial strain of the genus Roseburia.

In certain embodiments of the invention, the composition is for use intreating brain injury. The neuroprotective activity of the compositionsof the invention and their ability to reduce levels of histonedeacetylase activity (HDAC) may make them useful for treating braininjury. In preferred embodiments, the compositions of the invention arefor use in treating stroke, such as treating brain injury resulting froma stroke.

In developing the above invention, the inventors have identified andcharacterised a bacterial strain that is particularly useful fortherapy. The Roseburia intestinalis strain of the invention is shown tobe effective for treating cancer. The invention also providescompositions comprising such cells, or biologically pure cultures ofsuch cells. The invention also provides a cell of the Roseburiaintestinalis strain deposited under accession number NCIMB 43043, or aderivative thereof, for use in therapy, in particular for cancer.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1: Downregulation of IL-6 secretion

FIG. 2: Inhibition of α-synuclein induced NFκB promoter activation

FIG. 3: Inhibition of LPS induced NFκB promoter activation

FIG. 4: Change in antioxidant capacity

FIG. 5: Change in total anti-oxidant capacity (lipid oxidation)

FIG. 6: Change in histone deacetylatase (HDAC) activity

FIG. 7: Level of Indole production

FIG. 8: Level of Kyrunenine production

FIG. 9: Downregulation of IL-6 secretion by MRX001

FIGS. 10A-10B: Downregulation of IL-6 secretion by strain A: Inhibitionof IL-6 secretion in U373 (FIG. 10A); Inhibition of IL-6 secretion inU373 (FIG. 10B)

FIGS. 11A-11B: FIG. 11A) Inhibition of α-synuclein induced NFκB promoteractivation and FIG. 11B) Inhibition of LPS induced NFκB promoteractivation by strain A

FIGS. 12A-12B: Downregulation of IL-6 secretion by strain B: Inhibitionof IL-6 secretion in U373 (FIG. 12A); Inhibition of IL-6 secretion inU373 (FIG. 12B)

FIGS. 13A-13B: FIG. 13A) Inhibition of α-synuclein induced NFκB promoteractivation and FIG. 13B) Inhibition of LPS induced NFκB promoteractivation by strain B

DISCLOSURE OF THE INVENTION Bacterial Strains

The compositions of the invention comprise a bacterial strain of thegenus Roseburia. The examples demonstrate that bacteria of this genusare useful for treating or preventing neurodegenerative disorders. Thepreferred bacterial strains are of the species Roseburia hominis,Roseburia faecis and Roseburia intestinalis.

Examples of Roseburia species for use in the invention include Roseburiahominis, Roseburia cecicola, Roseburia faecis, Roseburia intestinalis,and Roseburia inulinivorans. Roseburia bacteria are slightly curvedrod-shaped cells that are strictly anaerobic and indigenous to themammalian intestine. They are of the phylogenetic cluster XIVa withinthe Firmicutes phylum. The bacteria are butyrate-producing and areactively motile through multiple flagella present along the concave sideand in a cluster at one end [23]. Roseburia hominis and Roseburiaintestinalis are recently described examples.

An example of Roseburia hominis is the strain deposited under the termsof the Budapest Treaty at National Collections of Industrial, Food andMarine Bacteria (NCIMB) at NCIMB Ltd, Ferguson Building, CraibstoneEstate, Bucksburn, Aberdeen, UK, AB21 9YA, on 21 Oct. 2004 by the RowettResearch Institute under the accession number NCIMB 14029^(T) Roseburiahominis A2-183^(T) (DSM=16839^(T)). Other exemplary Roseburia hominisstrains are described in [24]. GenBank/EMBL/DDBJ accession numbers forthe 16S rRNA gene sequence of strains of Roseburia hominis are AY804148and AJ270482 (disclosed herein as SEQ ID NO:1 and SEQ ID NO:2).

An example of Roseburia intestinalis is the strain deposited under theaccession number NCIMB 13810 Roseburia intestinalis L1-82^(T)(DSM=14610^(T)). Another example is the Roseburia intestinalis strain asdescribed in [24]. Reference [24] also describes exemplary Roseburiafaecis and Roseburia inulinivorans strains.

The Roseburia hominis bacterium deposited under accession number NCIMB42383 was tested in the Examples and is also referred to herein asstrain 433. A 16S rRNA sequence for the 433 strain that was tested isprovided in SEQ ID NO:3. Strain 433 was deposited with the internationaldepositary authority NCIMB, Ltd. (Ferguson Building, Aberdeen, AB21 9YA,Scotland) by GT Biologics Ltd. (Life Sciences Innovation Building,Aberdeen, AB25 2ZS, Scotland) on 12th March 2015 as “Roseburia hominis433” and was assigned accession number NCIMB 42383. GT Biologics Ltd.subsequently changed its name to 4D Pharma Research Limited.

WO 2016/203221 describes administration of strain 433 to mice and showsthat it can affect disease processes outside of the gut (such as asthmaand arthritis). Strain 433 also affects disease processes outside of thegut in the treatment of neurodegenerative disorders described herein.

A genome sequence for strain 433 is provided in SEQ ID NO:4. Thissequence was generated using the PacBio RS II platform.

The Roseburia intestinalis bacterium deposited under accession numberNCIMB 43043 was tested in the Examples and is also referred to herein asstrain A. A 16S rRNA sequence for strain A that was tested is providedin SEQ ID NO:5. Strain A was deposited with the international depositaryauthority NCIMB, Ltd. (Ferguson Building, Aberdeen, AB21 9YA, Scotland)by 4D Pharma Research Limited (Life Sciences Innovation Building,Aberdeen, AB25 2ZS, Scotland) on 3 May 2018 as “Roseburia intestinalis”and was assigned accession number NCIMB 43043.

The examples also describe a Roseburia faecis bacterium that was testedand referred to as strain B. A 16S rRNA sequence for strain B that wastested is provided in SEQ ID NO:6.

In certain embodiments, the bacterial strain for use in the invention isRoseburia hominis. In certain embodiments, the bacterial strain for usein the invention is Roseburia intestinalis. In certain embodiments, thebacterial strain for use in the invention is Roseburia faecis.

Bacterial strains closely related to the strain tested in the examplesare also expected to be effective for treating or preventingneurodegenerative disease diseases and conditions mediated by IL-17 orthe Th17 pathway. In certain embodiments, the bacterial strain for usein the invention has a 16S rRNA sequence that is at least 95%, 96%, 97%,98%, 99%, 99.5% or 99.9% identical to the 16S rRNA sequence of abacterial strain of Roseburia hominis. Preferably, the bacterial strainfor use in the invention has a 16S rRNA sequence that is at least 95%,96%, 97%, 98%, 99%, 99.5% or 99.9% identical to SEQ ID NO:1, 2 or 3.Preferably, the sequence identity is to SEQ ID NO:3. Preferably, thebacterial strain for use in the invention has the 16S rRNA sequencerepresented by SEQ ID NO:3.

In other embodiments, the bacterial strain for use in the invention hasa 16S rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or99.9% identical to the 16S rRNA sequence of a bacterial strain ofRoseburia intestinalis. Preferably, the bacterial strain for use in theinvention has a 16S rRNA sequence that is at least 95%, 96%, 97%, 98%,99%, 99.5% or 99.9% identical to SEQ ID NO:5. Preferably, the bacterialstrain for use in the invention has the 16S rRNA sequence represented bySEQ ID NO:5.

In other embodiments, the bacterial strain for use in the invention hasa 16S rRNA sequence that is at least 95%, 96%, 97%, 98%, 99%, 99.5% or99.9% identical to the 16S rRNA sequence of a bacterial strain ofRoseburia faecis. Preferably, the bacterial strain for use in theinvention has a 16S rRNA sequence that is at least 95%, 96%, 97%, 98%,99%, 99.5% or 99.9% identical to SEQ ID NO:6. Preferably, the bacterialstrain for use in the invention has the 16S rRNA sequence represented bySEQ ID NO:6.

Bacterial strains that are biotypes of the bacterium deposited underaccession number 42383 or NCIMB 43043 are also expected to be effectivefor treating or preventing neurodegenerative disorders. A biotype is aclosely related strain that has the same or very similar physiologicaland biochemical characteristics.

Strains that are biotypes of the bacterium deposited under accessionnumber NCIMB 42383 or NCIMB 43043 and that are suitable for use in theinvention may be identified by sequencing other nucleotide sequences forthe bacterium deposited under accession number NCIMB 42383 or NCIMB43043. For example, substantially the whole genome may be sequenced anda biotype strain for use in the invention may have at least 95%, 96%,97%, 98%, 99%, 99.5% or 99.9% sequence identity across at least 80% ofits whole genome (e.g. across at least 85%, 90%, 95% or 99%, or acrossits whole genome). Other suitable sequences for use in identifyingbiotype strains may include hsp60 or repetitive sequences such as BOX,ERIC, (GTG)₅ (SEQ ID NO: 7), or REP or [25]. Biotype strains may havesequences with at least 95%, 96%, 97%, 98%, 99%, 99.5% or 99.9% sequenceidentity to the corresponding sequence of the bacterium deposited underaccession number NCIMB 42383 or NCIMB 43043.

In certain embodiments, the bacterial strain for use in the inventionhas a genome with sequence identity to SEQ ID NO:4. In preferredembodiments, the bacterial strain for use in the invention has a genomewith at least 90% sequence identity (e.g. at least 92%, 94%, 95%, 96%,97%, 98%, 99% or 100% sequence identity) to SEQ ID NO:4 across at least60% (e.g. at least 65%, 70%, 75%, 80%, 85%, 95%, 96%, 97%, 98%, 99% or100%) of SEQ ID NO:4. For example, the bacterial strain for use in theinvention may have a genome with at least 90% sequence identity to SEQID NO:4 across 70% of SEQ ID NO:4, or at least 90% sequence identity toSEQ ID NO:4 across 80% of SEQ ID NO:4, or at least 90% sequence identityto SEQ ID NO:4 across 90% of SEQ ID NO:4, or at least 90% sequenceidentity to SEQ ID NO:4 across 100% of SEQ ID NO:4, or at least 95%sequence identity to SEQ ID NO:4 across 70% of SEQ ID NO:4, or at least95% sequence identity to SEQ ID NO:4 across 80% of SEQ ID NO:4, or atleast 95% sequence identity to SEQ ID NO:4 across 90% of SEQ ID NO:4, orat least 95% sequence identity to SEQ ID NO:4 across 100% of SEQ IDNO:4, or at least 98% sequence identity to SEQ ID NO:4 across 70% of SEQID NO:4, or at least 98% sequence identity to SEQ ID NO:4 across 80% ofSEQ ID NO:4, or at least 98% sequence identity to SEQ ID NO:4 across 90%of SEQ ID NO:4, or at least 98% sequence identity to SEQ ID NO:4 across100% of SEQ ID NO:4.

Alternatively, strains that are biotypes of the bacterium depositedunder accession number NCIMB 42383 or NCIMB 43043 and that are suitablefor use in the invention may be identified by using the accession numberNCIMB 42383 or NCIMB 43043 deposit and restriction fragment analysisand/or PCR analysis, for example by using fluorescent amplified fragmentlength polymorphism (FAFLP) and repetitive DNA element (rep)-PCRfingerprinting, or protein profiling, or partial 16S or 23S rDNAsequencing. In preferred embodiments, such techniques may be used toidentify other Roseburia hominis, Roseburia faecis and Roseburiaintestinalis strains.

In certain embodiments, strains that are biotypes of the bacteriumdeposited under accession number NCIMB 42383 or NCIMB 43043 and that aresuitable for use in the invention are strains that provide the samepattern as the bacterium deposited under accession number NCIMB 42383 orNCIMB 43043 when analysed by amplified ribosomal DNA restrictionanalysis (ARDRA), for example when using Sau3AI restriction enzyme (forexemplary methods and guidance see, for example,[25]). Alternatively,biotype strains are identified as strains that have the samecarbohydrate fermentation patterns as the bacterium deposited underaccession number NCIMB 42383 or NCIMB 43043.

Other Roseburia strains that are useful in the compositions and methodsof the invention, such as biotypes of the bacteria deposited underaccession number NCIMB 42383 or NCIMB 43043, may be identified using anyappropriate method or strategy, including the assays described in theexamples. For instance, strains for use in the invention may beidentified by culturing with neuroblastoma cells and then assessingcytokine levels and levels of neuroprotection or neuroproliferation. Inparticular, bacterial strains that have similar growth patterns,metabolic type and/or surface antigens to the bacterium deposited underaccession number NCIMB 42383 or NCIMB 43043 may be useful in theinvention. A useful strain will have comparable immune modulatoryactivity to the NCIMB 42383 or NCIMB 43043 strain. In particular, abiotype strain will elicit comparable effects on the neurodegenerativedisease models and comparable effects on cytokine levels to the effectsshown in the Examples, which may be identified by using the culturingand administration protocols described in the Examples.

A particularly preferred strain of the invention is the Roseburiahominis strain deposited under accession number NCIMB 42383. This is theexemplary 433 strain tested in the examples and shown to be effectivefor treating disease. Therefore, the invention provides a cell, such asan isolated cell, of the Roseburia hominis strain deposited underaccession number NCIMB 42383, or a derivative thereof. The inventionalso provides a composition comprising a cell of the Roseburia hominisstrain deposited under accession number NCIMB 42383, or a derivativethereof. The invention also provides a biologically pure culture of theRoseburia hominis strain deposited under accession number NCIMB 42383.The invention also provides a cell of the Roseburia hominis straindeposited under accession number NCIMB 42383, or a derivative thereof,for use in therapy, in particular for the diseases described herein.

A particularly preferred strain of the invention is the Roseburiaintestinalis strain deposited under accession number NCIMB 43043. Thisis the exemplary strain A tested in the examples and shown to beeffective for treating disease. Therefore, the invention provides acell, such as an isolated cell, of the Roseburia intestinalis straindeposited under accession number NCIMB 43043, or a derivative thereof.The invention also provides a composition comprising a cell of theRoseburia intestinalis strain deposited under accession number NCIMB43043, or a derivative thereof. The invention also provides abiologically pure culture of the Roseburia intestinalis strain depositedunder accession number NCIMB 43043. The invention also provides a cellof the Roseburia intestinalis strain deposited under accession numberNCIMB 43043, or a derivative thereof, for use in therapy, in particularfor the diseases described herein.

A derivative of the strain deposited under accession number NCIMB 42383or NCIMB 43043 may be a daughter strain (progeny) or a strain cultured(subcloned) from the original. A derivative of a strain of the inventionmay be modified, for example at the genetic level, without ablating thebiological activity. In particular, a derivative strain of the inventionis therapeutically active. A derivative strain will have comparableimmune modulatory activity to the original NCIMB 42383 or NCIMB 43043strain. In particular, a derivative strain will elicit comparableeffects on the neurodegenerative disease models and comparable effectson cytokine levels to the effects shown in the Examples, which may beidentified by using the culturing and administration protocols describedin the Examples. A derivative of the NCIMB 42383 or NCIMB 43043 strainwill generally be a biotype of the NCIMB 42383 or NCIMB 43043 strain.

References to cells of the Roseburia hominis strain deposited underaccession number NCIMB 42383 encompass any cells that have the samesafety and therapeutic efficacy characteristics as the strains depositedunder accession number NCIMB 42383, and such cells are encompassed bythe invention.

References to cells of the Roseburia intestinalis strain deposited underaccession number NCIMB 43043 encompass any cells that have the samesafety and therapeutic efficacy characteristics as the strains depositedunder accession number NCIMB 43043, and such cells are encompassed bythe invention.

In preferred embodiments, the bacterial strains in the compositions ofthe invention are viable and capable of partially or totally colonisingthe intestine.

The invention further provides a cell, such as an isolated cell, of theEnterococcus gallinarum strain deposited under accession number NCIMB42761, or a derivative thereof. The invention also provides acomposition comprising a cell of the Roseburia intestinalis straindeposited under accession number NCIMB 42761, or a derivative thereof.The invention also provides a biologically pure culture of the Roseburiaintestinalis strain deposited under accession number NCIMB 43043. Theinvention also provides a cell of the Roseburia intestinalis straindeposited under accession number NCIMB 43043, or a derivative thereof,for use in therapy, in particular for the diseases described herein. Aderivative of the strain deposited under accession number NCIMB 43043may be a daughter strain (progeny) or a strain cultured (subcloned) fromthe original.

A derivative of a strain of the invention may be modified, for exampleat the genetic level, without ablating the biological activity. Inparticular, a derivative strain of the invention is therapeuticallyactive. A derivative strain will have comparable immune modulatoryactivity to the original NCIMB 42761 strain. In particular, a derivativestrain will elicit comparable effects on the neurodegenerative diseasemodels to the effects shown in the Examples, which may be identified byusing the culturing and administration protocols described in theExamples. A derivative of the NCIMB 43043 strain will generally be abiotype of the NCIMB 43043 strain.

References to cells of the Roseburia intestinalis strain deposited underaccession number NCIMB 43043 encompass any cells that have the samesafety and therapeutic efficacy characteristics as the strain depositedunder accession number NCIMB 43043, and such cells are encompassed bythe invention. Thus, in some embodiments, reference to cells of theRoseburia intestinalis strain deposited under accession number NCIMB43043 refers only to the strain A deposited under NCIMB 43043 and doesnot refer to a bacterial strain that was not deposited under NCIMB43043.

In preferred embodiments, the bacterial strain in the compositions ofthe invention is viable and capable of partially or totally colonisingthe intestine.

Therapeutic Uses

As demonstrated in the examples, the bacterial compositions of theinvention are effective for treating neurodegenerative disorders. Inparticular, treatment with compositions of the invention increaseneuroproliferation and act as a neuroprotectant against agents thatdestroy dopaminergic neurons. Therefore, the compositions of theinvention may be useful for treating or preventing neurodegenerativedisorders that are the result of neuron death.

Compositions of the invention can decrease the activation of the NFκBpromoter, which activates cytokine production, for example IL-1β, IL-1α,IL-18, TNFα and IL-6. Treating cells with mutant α-synuclein is a modelfor familial Parkinson's. A point mutation at position 53 from adenineto threonine leads to α-synuclein mis-folding. The incorrectly foldedα-synuclein subsequently aggregates into insoluble fibrils which formLewy bodies. Therefore, the compositions of the invention may be usefulfor treating or preventing neurodegenerative disorders that are theresult of neuroinflammation, protein misfolding and/or environmentalexposure. Compositions of the invention can be used for treatment offamilial Parkinson's. Activation of the NFκB promoter is mediatedthrough the TLR4 ligand. TL4 is known to mediate cell death in the mousemodel MPTP, which simulates Parkinson's disease. Compositions of theinvention can be used to inhibit the ability of TLR4 signalling toactivate the NFκB promoter. Of particular relevance for PD, both TLR2and TLR4 were found to be upregulated in brains of PD patients [26].Moreover α-syn has been described as a ligand for TLR2 [27] and we havedemonstrated that α-syn is also a ligand for TLR4 using HEK-TLR4 cells.

Compositions of the invention decrease the secretion of pro-inflammatorycytokines such as IL-6, which can be induced by lipopolysaccharide(LPS). Treatment of cells with LPS simulates Parkinson's caused byenvironmental factors. Compositions of the invention can be used todecrease IL-6 secretion. Compositions of the invention can be used fortreatment of environmental Parkinson's.

Examples of neurodegenerative diseases to be treated by compositions ofthe invention include: Parkinson's disease, including progressivesupranuclear palsy, progressive supranuclear palsy,Steele-Richardson-Olszewski syndrome, normal pressure hydrocephalus,vascular or arteriosclerotic parkinsonism and drug-induced parkinsonism;Alzheimer's disease, including Benson's syndrome; multiple sclerosis;Huntington's disease; amyotrophic lateral sclerosis; Lou Gehrig'sdisease; motor neurone disease; prion disease; spinocerebellar ataxia;spinal muscular atrophy; dementia, including Lewy body, vascular andfrontotemporal dementia; primary progressive aphasia; mild cognitiveimpairment; HIV-related cognitive impairment and corticobasaldegeneration. A further neurodegenerative diseases to be treated bycompositions of the invention is progressive inflammatory neuropathy.

In certain embodiments, the compositions of the invention can beeffective for treating neurodegenerative disorders that occur in elderlypatients. The examples show that compositions of the invention can treatParkinson's disease which is predominantly diagnosed in patients agedover 65 years old. In preferred embodiments, the compositions of theinvention are for treating patients 65 years or older. In other certainembodiments, the patients are between 40 to 65 years old. In otherembodiments, the patients are older than 40 years. In certainembodiments, the compositions of the invention are for use in treating adisease associated with old age, for example, a disease diagnosed after50 years of age.

In certain embodiments, the compositions of the invention are for use intreating a neurodegenerative disorder mediated or characterised by theaccumulation of protein, in particular mis-folded protein.

In certain embodiments, the compositions of the invention are for use intreating a neurodegenerative disorder associated with grey matterneuronal loss. In certain embodiments, the compositions of the inventionare for treating a neurodegenerative disorder that is not associatedwith white matter lesions.

In certain embodiments, the compositions of the invention are for use intreating a neurodegenerative disorder associated with permanentsymptoms.

In certain embodiments, the compositions of the invention are for use intreating a neurodegenerative disorder that is not an auto-immunedisorder. In certain embodiments, the compositions of the invention arefor use in treating a neurodegenerative disorder that is not multiplesclerosis.

In certain embodiments, the compositions of the invention are for use inreducing neuron death, in particular, in the treatment ofneurodegenerative disorders. In certain embodiments, the compositions ofthe invention are for use in protecting neurons, in particular in thetreatment of neurodegenerative disorders.

The neuroprotective properties of the compositions of the invention, asshown in the examples, mean that the compositions may be particularlyeffective for preventing or delaying onset or progression ofneurodegenerative disorders. In certain embodiments, the compositions ofthe invention are for use in delaying onset or progression of aneurodegenerative disorders.

Modulation of the Microbiota-Gut-Brain Axis

Communication between the gut and the brain (the microbiota-gut-brainaxis) occurs via a bidirectional neurohumoral communication system.Recent evidence shows that the microbiota that resides in the gut canmodulate brain development and produce behavioural phenotypes via themicrobiota-gut-brain axis. Indeed, a number of reviews suggest a role ofthe microbiota-gut-brain axis in maintaining central nervous systemfunctionality and implicate dysfunction of the microbiota-gut-brain axisin the development of central nervous system disorders and conditions[17],[20],[28].

The bidirectional communication between the brain and the gut (i.e.the-gut-brain axis) includes the central nervous system, neuroendocrineand neuroimmune systems, including the hypothalamus-pituitary-adrenal(HPA) axis, sympathetic and parasympathetic arms of the autonomicnervous system (ANS), including the enteric nervous system (ENS) and thevagus nerve, and the gut microbiota.

As demonstrated in the examples, the compositions of the presentinvention can modulate the microbiota-gut-brain axis and reduce celldeath associated with neurodegenerative disorders. Accordingly, thecompositions of the invention may be useful for treating or preventingneurodegenerative disorders, in particular those disorders andconditions associated with dysfunction of the microbiota-gut-brain axis.

In particular embodiments, the compositions of the invention may beuseful for treating or preventing a disease or condition selected fromthe group consisting of: Parkinson's disease, including progressivesupranuclear palsy, progressive supranuclear palsy,Steele-Richardson-Olszewski syndrome, normal pressure hydrocephalus,vascular or arteriosclerotic parkinsonism and drug-induced parkinsonism;Alzheimer's disease, including Benson's syndrome; multiple sclerosis;Huntington's disease; amyotrophic lateral sclerosis; Lou Gehrig'sdisease; motor neurone disease; prion disease; spinocerebellar ataxia;spinal muscular atrophy; dementia; including Lewy body; vascular andfrontotemporal dementia; primary progressive aphasia; mild cognitiveimpairment; HIV-related cognitive impairment and corticobasaldegeneration.

The compositions of the invention may be particularly useful fortreating or preventing chronic disease, treating or preventing diseasein patients that have not responded to other therapies (such astreatment with Levodopa, dopamine agonists, MAO-B inhibitors, COMTinhibitors, Glutamate antagonists, and/or anticholinergics), and/ortreating or preventing the tissue damage and symptoms associated withdysfunction of the microbiota-gut-brain axis.

In certain embodiments, the compositions of the invention modulate theCNS. In some embodiments, the compositions of the invention modulate theautonomic nervous system (ANS). In some embodiments, the compositions ofthe invention modulate the enteric nervous system (ENS). In someembodiments, the compositions of the invention modulate thehypothalamic, pituitary, adrenal (HPA) axis. In some embodiments, thecompositions of the invention modulate the neuroendocrine pathway. Insome embodiments, the compositions of the invention modulate theneuroimmune pathway. In some embodiments, the compositions of theinvention modulate the CNS, the ANS, the ENS, the HPA axis and/or theneuroendocrine and neuroimmune pathways. In certain embodiments, thecompositions of the invention module the levels of commensal metabolitesand/or the gastrointestinal permeability of a subject. In certainembodiments, the compositions of the invention modulate the dopaminergicsystem.

The signalling of the microbiota-gut-brain axis is modulated by neuralsystems. Accordingly, in some embodiments, the compositions of theinvention modulate signalling in neural systems. In certain embodiments,the compositions of the invention modulate the signalling of the centralnervous system. In some embodiments, the compositions of the inventionmodulate signalling in sensory neurons. In other embodiments, thecompositions of the invention modulate signalling in motor neurons. Insome embodiments, the compositions of the invention modulate thesignalling in the ANS. In some embodiments, the ANS is theparasympathetic nervous system. In preferred embodiments, thecompositions of the invention modulate the signalling of the vagusnerve. In other embodiments, the ANS is the sympathetic nervous system.In other embodiments, the compositions of the invention modulate thesignalling in the enteric nervous system. In certain embodiments, thesignalling of ANS and ENS neurons responds directly to luminal contentsof the gastrointestinal tract. In other embodiments, the signalling ofANS and ENS neurons responds indirectly to neurochemicals produced byluminal bacteria. In other embodiments, the signalling of ANS and ENSneurons responds to neurochemicals produced by luminal bacteria orenteroendocrine cells. In certain preferred embodiments, the neurons ofthe ENS activate vagal afferents that influence the functions of theCNS. In some embodiments, the compositions of the invention regulate theactivity of enterochromaffin cells.

Neurodegenerative Diseases

Tauopathies are neurodegenerative diseases associated with thepathological aggregation of tau protein in neurofibrillary orgliofibrillary tangles in the human brain. Alzheimer's disease is anexample of a tauopathology. Synucleinopathies (also calledα-Synucleinopathies) are neurodegenerative diseases characterised by theabnormal accumulation of aggregates of α-synuclein in neurons, nervefibres or glial cells. Parkinson's disease is an example of asynucleinopathology.

There is clinical and pathological overlap between these twopathologies. Parkinson's disease patients frequently have dementia andAlzheimer's disease patients often manifest parkinsonism [29]. Forexample, progressive supranuclear palsy (also known asSteele-Richardson-Olszewski syndrome) has a tauopathology, but alsoleads to prominent parkinsonism [30]. Mutations in LRRK2 known to causeparkinsonism are associated with the accumulation of synuclein, tau,neither, or both proteins [31].

Lewy body disease (LBD) is a neurodegenerative disease that is one ofthe most common causes of dementia in the elderly. LBD exemplifies theexistence of a continuum between tau- and synuclein-pathologies. LBDshares clinical and pathological features with Parkinson disease,Parkinson disease dementia and Alzheimer disease [29].

In particular embodiments, the compositions of the invention may beuseful for treating or preventing tauopathies and/or synucleinopathies.In particular embodiments, the compositions of the invention may beuseful for treating or preventing tauopathies. In particularembodiments, the compositions of the invention may be useful fortreating or preventing synucleinopathies. In certain embodiments, thecompositions of the invention may be useful for treating or preventing adisease or condition selected from the group consisting of: Parkinson'sdisease, including progressive supranuclear palsy, progressivesupranuclear palsy, Steele-Richardson-Olszewski syndrome, normalpressure hydrocephalus, vascular or arteriosclerotic parkinsonism anddrug-induced parkinsonism; Alzheimer's disease, including Benson'ssyndrome; and dementia; including Lewy body; vascular and frontotemporaldementia.

In preferred embodiments, the compositions of the invention may beuseful for treating or preventing Parkinson's disease, includingprogressive supranuclear palsy, progressive supranuclear palsy,Steele-Richardson-Olszewski syndrome, normal pressure hydrocephalus,vascular or arteriosclerotic parkinsonism and drug-induced parkinsonism.In preferred embodiments, the compositions of the invention may beuseful for treating or preventing Alzheimer's disease, includingBenson's syndrome. In further preferred embodiments, the compositions ofthe invention may be useful for treating or preventing dementia;including Lewy body; vascular and frontotemporal dementia.

Parkinson's Disease

Parkinson's disease is a common neurodegenerative diseaseneuropathologically characterised by degeneration of heterogeneouspopulations of neural cells (dopamine-producing cells). The clinicaldiagnosis of Parkinson's disease requires bradykinesia and at least oneof the following core symptoms: resting tremor; muscle rigidity andpostural reflex impairment. Other signs and symptoms that may be presentor develop during the progression of the disease are autonomicdisturbances (sialorrhoea, seborrhoea, constipation, micturitiondisturbances, sexual functioning, orthostatic hypotension,hyperhydrosis), sleep disturbances and disturbances in the sense ofsmell or sense of temperature. Parkinson's disease is aneurodegenerative diseases that may develop or persist due todysfunction of the microbiota-gut-brain axis. Therefore, in preferredembodiments, the compositions of the invention are for use in treatingor preventing Parkinson's disease in a subject.

In further preferred embodiments, the invention provides a compositioncomprising a bacterial strain of the genus Roseburia, for use in amethod of treating or preventing Parkinson's disease. Compositionscomprising a bacterial strain of the genus Roseburia may improve motorand cognitive functions in models of Parkinson's disease. Treatment withRoseburia strains may modulate signalling in the central, autonomic andenteric nervous systems; may modulate the activity of the HPA axispathway; may modulate neuroendocrine and/or neuroimmune pathways; andmay modulate the levels of commensal metabolites, inflammatory markersand/or gastrointestinal permeability of a subject, all of which areimplicated in the neuropathology of Parkinson's disease. In preferredembodiments, the invention provides a composition comprising a bacterialstrain of the species Roseburia hominis for use in a method of treatingor preventing Parkinson's disease. Compositions using Roseburia hominismay be particularly effective for treating Parkinson's disease.

In preferred embodiments, the compositions of the invention prevent,reduce or alleviate one or more of the symptoms of Parkinson's diseasein a subject. In preferred embodiments, the compositions of theinvention prevent, reduce or alleviate one or more core symptoms ofParkinson's disease in a subject. In certain embodiments, thecompositions of the invention prevent, reduce or alleviate bradykinesiain a subject. In certain embodiments, the compositions of the inventionprevent, reduce or alleviate resting tremor; muscle rigidity and/orpostural reflex impairment in a subject. In certain embodiments, thecompositions of the invention prevent, reduce or alleviate one or moresymptoms associated with Parkinson's disease progression selected fromautonomic disturbances (sialorrhoea, seborrhoea, constipation,micturition disturbances, sexual functioning, orthostatic hypotension,hyperhydrosis), sleep disturbances and disturbances in the sense ofsmell or sense of temperature.

In preferred embodiments, the compositions of the invention prevent,reduce or alleviate depressive symptoms comorbid with Parkinson'sdisease. In certain embodiments, the compositions of the inventionimprove verbal memory and/or executive functions. In certainembodiments, the compositions of the invention improve attention,working memory, verbal fluency and/or anxiety.

In other preferred embodiments, the compositions of the inventionprevent, reduce or alleviate cognitive dysfunctions comorbid withParkinson's disease.

In certain embodiments, the compositions of the invention prevent,reduce or alleviate Parkinson's disease progression. In certainembodiments, the compositions of the invention prevent, reduce oralleviate later motor complications. In certain embodiments, thecompositions of the invention prevent, reduce or alleviate late motorfluctuations. In certain embodiments, the compositions of the inventionprevent, reduce or alleviate neuronal loss. In certain embodiments, thecompositions of the invention improve symptoms of Parkinson's diseasedementia (PDD). In certain embodiments, the compositions of theinvention prevent, reduce or alleviate impairment of executive function,attention and/or working memory. In certain embodiments, thecompositions of the invention improve dopaminergic neurotransmission. Incertain embodiments, the compositions of the invention prevent, reduceor alleviate impaired dopaminergic neurotransmission.

In some embodiments, the compositions of the invention improve thesymptoms of Parkinson's disease according to a symptomatic or diagnosticscale. In certain embodiments, the tests for assessing symptomaticimprovement of motor function in Parkinson's disease is the UnifiedParkinson's Disease Rating Scale. In particular, UPDRS II considers theactivity of daily life and UPDRS III considers motor-examination.

In some embodiments, the compositions of the invention improve thesymptoms associated the PDD according to a symptomatic or diagnostictest and/or scale. In certain embodiments, the test or scale is selectedfrom the Hopkins Verbal Learning Test—Revised (HVLT-R); the Delis-KaplanExecutive Function System (D-KEFS) Color-Word Interference Test; theHamilton Depression Rating Scale (HAM-D 17; depression); the HamiltonAnxiety Rating Scale (HAM-A; anxiety) and the Unified Parkinson'sDisease Rating Scale (UPDRS; PD symptom severity).

In some embodiments, the compositions of the invention improve theClinical Global Impression—Global Improvement (CGI-I) scale forassessing psychiatric and neurological disorders. In some embodiments,the compositions of the invention display a positive effect on globalsocial and occupational impairment of the subject with Parkinson'sdisease.

Alzheimer's Disease and Dementia

In DSM-5, the term dementia was replaced with the terms majorneurocognitive disorder and mild neurocognitive disorder. Neurocognitivedisorder is a heterogeneous class of psychiatric diseases. The mostcommon neurocognitive disorder is Alzheimer's disease, followed byvascular dementias or mixed forms of the two. Other forms ofneurodegenerative disorders (e.g. Lewy body disease, frontotemporaldementia, Parkinson's dementia, Creutzfeldt-Jakob disease, Huntington'sdisease, and Wernicke-Korsakoff syndrome) are accompanied by dementia.

Alzheimer's disease and dementia are also characterised by neuronalloss, so the neuroprotective and neuroproliferative effects shown in theexamples for the compositions of the invention indicate that they may beuseful for treating or preventing these conditions.

The symptomatic criteria for dementia under DSM-5 are evidence ofsignificant cognitive decline from a previous level of performance inone or more cognitive domains selected from: learning and memory;language; executive function; complex attention; perceptual-motor andsocial cognition. The cognitive deficits must interfere withindependence in everyday activities. In addition, the cognitive deficitsdo not occur exclusively in the context of a delirium and are not betterexplained by another mental disorder (for example MDD or schizophrenia).

In addition to the primary symptom, subjects with neurodegenerativedisorders display behavioural and psychiatric symptoms includingagitation, aggression, depression, anxiety, apathy, psychosis andsleep-wake cycle disturbances.

Neurodegenerative disorders may develop or persist due to dysfunction ofthe microbiota-gut-brain axis. Therefore, in preferred embodiments, thecompositions of the invention are for use in treating or preventingneurodegenerative disorders in a subject. In preferred embodiments, theneurodegenerative disorder is Alzheimer's disease. In other embodiments,the neurodegenerative disorder is selected from vascular dementias;mixed form Alzheimer's disease and vascular dementia; Lewy body disease;frontotemporal dementia; Parkinson's dementia; Creutzfeldt-Jakobdisease; Huntington's disease; and Wernicke-Korsakoff syndrome.

In preferred embodiments, the compositions of the invention prevent,reduce or alleviate one or more of the symptoms of neurodegenerativedisorders in a subject. In certain embodiments, the compositions of theinvention prevent, reduce or alleviate the occurrence of cognitivedecline in a subject. In certain embodiments, the compositions of theinvention improve the level of performance of a subject withneurodegenerative disorders in one or more cognitive domains selectedfrom: learning and memory; language; executive function; complexattention; perceptual-motor and social cognition. In some embodiments,the compositions of the invention prevent, reduce or alleviate theoccurrence of one or more behavioural and psychiatric symptomsassociated with neurodegenerative disorders selected from agitation,aggression, depression, anxiety, apathy, psychosis and sleep-wake cycledisturbances.

In certain embodiments, the compositions of the invention prevent,reduce or alleviate symptomatic disease by intervention in suspectedpathogenic mechanisms at a preclinical stage. In certain embodiments,the compositions of the invention improve disease modification, withslowing or arrest of symptom progression. In some embodiments, theslowing or arrest of symptom progression correlates with evidence indelaying the underlying neuropathological process. In preferredembodiments, the compositions of the invention improve symptoms ofneurodegenerative disorders comprising enhanced cognitive and functionalimprovement. In preferred embodiments, the compositions of the inventionimprove the behavioural and psychiatric symptoms of dementia (BPSD). Inpreferred embodiments, the compositions of the invention improve theability of a subject with neurodegenerative disorder to undertakeeveryday activities.

In preferred embodiments, the compositions of the invention improve bothcognition and functioning in a subject with Alzheimer's disease. In someembodiments, the composition of the invention improves the cognitiveendpoint in a subject with Alzheimer's disease. In some embodiments, thecompositions of the invention improve the functional endpoint in asubject with Alzheimer's disease. In preferred embodiments, thecompositions of the invention improve the cognitive and functionalendpoint in a subject with Alzheimer's disease. In yet further preferredembodiments, the compositions of the invention improve the overallclinical response (the global endpoint) in a subject with Alzheimer'sdisease.

In some embodiments, the compositions of the invention improve thesymptoms of neurodegenerative disorders according to a symptomatic ordiagnostic test. In certain embodiments, the tests for assessingsymptomatic improvement of Alzheimer's disease (and otherneurodegenerative disorders) are selected from objective cognitive,activities of daily living, global assessment of change, health relatedquality of life tests and tests assessing behavioural and psychiatricsymptoms of neurodegenerative disorders.

In certain embodiments, the objective cognitive tests for assessment ofsymptomatic improvement use the Alzheimer's disease Assessment Scalecognitive subscale (ADAS-cog) and the classic ADAS scale. In certainembodiments, symptomatic improvement of cognition is assessed using theNeurophysiological Test Battery for Use in Alzheimer's Disease (NTB).

In some embodiments, the global assessment of change test uses theClinical Global Impression—Global Improvement (CGI-I) scale forassessing psychiatric and neurological disorders. In some embodiments,the global scale is the Clinician's Interview Based Impression of Changeplus (CIBIC-plus). In some embodiments, the global scale is theAlzheimer's Disease Cooperative Study Unit Clinician's Global Impressionof Change (ADCS-CGIC).

In certain embodiments, the health related quality of life measures arethe Alzheimer's Disease-Related QOL (ADRQL) and the QOL-Alzheimer'sDisease (QOL-AD).

In certain embodiments, the tests assessing behavioural and psychiatricsymptoms of neurodegenerative disorders are selected from theBehavioural pathology in Alzheimer's Disease Rating Scale (BEHAVE-AD);the Behavioural Rating Scale for Dementia (BRSD); the NeuropsychiatricInventory (NPI); and the Cohen-Mansfield Agitation Inventory (CMAI).

In some embodiments, the compositions of the invention are particularlyeffective at preventing, reducing or alleviating neurodegenerativedisorders when used in combination with another therapy for treatingneurodegenerative disorders. In certain embodiments, such therapiesinclude acetylcholinesterase inhibitors including donepezil (Aricept®),galantamine (Razadyne®) and rivastigmine (Exelon®), and memantine.

Neurochemical Factors, Neuropeptides and Neurotransmitters and theMicrobiota-Gut-Brain Axis

As outlined above, the microbiota-gut-brain axis is modulated by anumber of different physiological systems. The microbiota-gut-brain axisis modulated by a number of signalling molecules. Alterations in thelevels of these signalling molecules results in neurodegenerativediseases. The experiments performed by the inventors indicate thatadministration of Roseburia species, and in particular Roseburiahominis, can modulate levels of indole and kynurenine. Dysregulation ofthese metabolites can lead to neurodegenerative diseases, such asParkinson's disease.

In certain embodiments, the compositions of the invention modulate thelevels of brain monoamines and metabolites thereof. In preferredembodiments the metabolite is kynurenine. In certain embodiments, thecompositions of the invention modulate kynurenine, which is the mainroute of tryptophan metabolism, which serves as a route to nicotinamideadenine dinucleotide (NAD+) production. Kynurenine can be metabolized toneuroactive compounds such as kynurenic acid (KYNA) and3-hydroxy-1-kynurenine (3-OH-1-KYN), and in further steps to quinolinicacid (QUIN). Dysregulation of the kynurenine pathway can lead toactivation of the immune system and the accumulation of potentiallyneurotoxic compounds. Alterations in the kynurenine metabolism may beinvolved in the development of Parkinson's diseases. Kynurenine levelshave been demonstrated to be decreased in the frontal cortex, putamenand substantia nigra pars compacta of patients with PD [32]. Therefore,in certain embodiments the compositions of the invention are for use inincreasing the levels of kynurenine in the treatment of Parkinson'sdisease.

In certain embodiments of the invention the compositions of theinvention can increase the levels of kynurenin. Increased levels ofkynurenine have been shown to attenuated MPP+-induced neuronal celldeath in vitro in a human dopaminergic neuroblastoma cell line [33]. Incertain embodiments kynurenine and kynurenic acid, can activate GI arylhydrocarbon receptor (Ahr) and GPR35 receptors. Activation of Ahrreceptor induces IL-22 production, which can inhibit local inflammation.Activation of GPR35 inducing the production of inositol triphosphate andCa2+ mobilization.

In certain embodiments, the compositions of the invention modulate thelevels of indole. In preferred embodiments the metabolite is kynurenine.In certain embodiments, the compositions of the invention modulatekynurenine, which is the main route of tryptophan metabolism.

The signalling of the microbiota-gut-brain axis is modulated by levelsof neurochemical factors, neuropeptides and neurotransmitters.Accordingly, in certain embodiments, the compositions of the inventionmodulates levels of neurochemical factors, neuropeptides andneurotransmitters. Accordingly, in certain preferred embodiments, thecompositions of the invention directly alter CNS biochemistry.

The signalling of the microbiota-gut-brain axis is modulated by levelsof γ-aminobutyric acid (GABA). Accordingly, in preferred embodiments,the compositions of the invention modulate the levels of GABA. GABA isan inhibitory neurotransmitter that reduces neuronal excitability. Incertain embodiments, the compositions of the invention increase thelevels of GABA. In certain embodiments, the compositions of theinvention decrease the levels of GABA. In certain embodiments, thecompositions of the invention alter GABAergic neurotransmission. Incertain embodiments, the compositions of the invention modulate thelevel of GABA transcription in different regions of the central nervoussystem. In certain embodiments, the commensal derived GABA crosses theblood-brain barrier and affects neurotransmission directly. In certainembodiments, the compositions of the invention lead to a reduction ofGABA in the hippocampus, amygdala and/or locus coeruleus. In certainembodiments, the compositions of the invention lead to an increase ofGABA in cortical regions.

Immune Response

The signalling of the microbiota-gut-brain axis is modulated byalterations in the immune response and inflammatory factors and markers.Accordingly, in certain embodiments, the compositions of the inventionmay modulate the immune response. In certain embodiments, thecompositions of the invention modulate the systemic levels ofcirculating neuroimmune signalling molecules. In certain preferredembodiments, the compositions of the invention modulate pro-inflammatorycytokine production and inflammation. In certain embodiments, thecompositions of the invention modulate the inflammatory state. Incertain embodiments, the compositions of the invention decrease IL-6production and secretion. In certain embodiments, the compositions ofthe invention decrease the activation of the NFκB promoter. In certainembodiments, the compositions of the invention are able to modulate theactivation of IL-6 production by the potent pro-inflammatory endotoxinlipopolysaccharide (LPS). In certain embodiments, the compositions ofthe invention are able to modulate the activation of the NFκB promoterby LPS and α-synuclein mutant proteins such as A53T. Increasedcirculating levels of cytokines are closely associated with variousneurodegenerative disorders, including Parkinson's, dementia andAlzheimer's. In certain embodiments, the compositions of the inventionare for use in reducing IL-6 levels and/or NFκB levels in the treatmentof a neurodegenerative disorder.

The signalling of the microbiota-gut-brain axis is modulated by levelsof commensal metabolites. Accordingly, in certain embodiments, thecompositions of the invention modulate the systemic levels of microbiotametabolites. In certain preferred embodiments, the compositions of theinvention modulate the level of short chain fatty acids (SCFAs). Incertain embodiments the level of SCFAs is increased or decreased. Insome embodiments, the SCFA is butyric acid (BA) (or butyrate). In someembodiments, the SCFA is propionic acid (PPA). In some embodiments, theSCFA is acetic acid. In certain embodiments, the compositions of theinvention modulate the ability of SCFAs to cross the blood-brainbarrier.

Histone acetylation and deacetylation are important epigeneticregulators of gene expression. An imbalance in histone acetylation anddeacetylation can result in apoptosis. Dysregulation of such histoneacetyltransferases has been implicated in the pathogenesis associatedwith age-associated neurodegenerative diseases, such as Parkinson'sdisease, Huntington's disease, Alzheimer's disease, amyotrophic lateralsclerosis and cognitive decline [34]. Accordingly, in certainembodiments, the compositions of the invention can modulate histonedeacetylatase activity. In certain embodiments, the compositions of theinvention can reduce histone deacetylatase activity. In certainembodiments, the compositions of the invention can reduce histoneacetylatase activity.

Patients with neurodegenerative diseases, including Parkinson's disease,Huntington's disease, Alzheimer's disease and amyotrophic lateralsclerosis, exhibit high levels of lipid peroxidation. Lipid arevulnerable to oxidation by reactive oxygen species, and the brain isrich in polyunsaturated fatty acids. Accordingly, in certainembodiments, the compositions of the invention can modulate lipidperoxidation. In certain embodiments, the compositions of the inventioncan reduce lipid peroxidation. Reducing the oxidative damage caused byreactive oxygen species can be used to target early the stagesneurodegenerative diseases. Accordingly, in certain embodiments, thecompositions of the invention are for use in treating early stageneurodegeneration. Also accordingly, in certain embodiments, thecompositions of the invention are for use in preventing the developmentof a neurodegenerative disorder. In such embodiments, the compositionsof the invention may be for use in a patient that has been identified asat risk of developing a neurodegenerative disorder.

The signalling of the microbiota-gut-brain axis is modulated by levelsof gastrointestinal permeability. Accordingly, in some embodiments, thecompositions of the invention alter the integrity of thegastrointestinal tract epithelium. In certain embodiments, thecompositions of the invention modulate the permeability of thegastrointestinal tract. In certain embodiments, the compositions of theinvention modulate the barrier function and integrity of thegastrointestinal tract. In certain embodiments, the compositions of theinvention modulate gastrointestinal tract motility. In certainembodiments, the compositions of the invention modulate thetranslocation of commensal metabolites and inflammatory signallingmolecules into the bloodstream from the gastrointestinal tract lumen.

The signalling of the microbiota-gut-brain axis is modulated bymicrobiome composition in the gastrointestinal tract. Accordingly, incertain embodiments, the compositions of the invention modulates themicrobiome composition of the gastrointestinal tract. In certainembodiments, the compositions of the invention prevents microbiomedysbiosis and associated increases in toxic metabolites (e.g. LPS). Incertain embodiments, the compositions of the invention modulate thelevels of Clostridium in the gastrointestinal tract. In preferredembodiments, the compositions of the invention reduce the level ofClostridium in the gastrointestinal tract. In certain embodiments, thecompositions of the invention reduce the levels of Campylobacter jejuni.In certain embodiments, the compositions of the invention modulate theproliferation of harmful anaerobic bacteria and the production ofneurotoxins produced by these bacteria. In certain embodiments, thecompositions of the invention modulate the microbiome levels ofLactobacillus and/or Bifidobacterium. In certain embodiments, thecompositions of the invention modulate the microbiome levels ofSutterella, Prevotella, Ruminococcucs genera and/or the Alcaligenaceaefamily. In certain embodiments, the compositions of the inventionincrease the level of Lactobacillus plantarum and/or Saccharomycesboulardii.

Brain Injury

The examples demonstrate that the compositions of the invention areneuroprotective and have HDAC inhibitory activity. HDAC2 is a crucialtarget for functional recovery from stroke [35] and HDAC inhibition canprevent white matter injury [36], so the compositions of the inventionmay be useful in the treatment of brain injury.

In certain embodiments, the compositions of the invention are for use intreating brain injury. In some embodiments, the brain injury is atraumatic brain injury. In some embodiments, the brain injury is anacquired brain injury. In some embodiments, the compositions of theinvention are for use in treating brain injury resulting from trauma. Insome embodiments, the compositions of the invention are for use intreating brain injury resulting from a tumour. In some embodiments, thecompositions of the invention are for use in treating brain injuryresulting from a stroke. In some embodiments, the compositions of theinvention are for use in treating brain injury resulting from a brainhaemorrhage. In some embodiments, the compositions of the invention arefor use in treating brain injury resulting from encephalitis. In someembodiments, the compositions of the invention are for use in treatingbrain injury resulting from cerebral hypoxia. In some embodiments, thecompositions of the invention are for use in treating brain injuryresulting from cerebral anoxia.

In preferred embodiments, the compositions of the invention are for usein treating stroke. The effects shown in the examples are particularlyrelevant to the treatment of stroke. Stroke occurs when blood flow to atleast a part of the brain is interrupted. Without an adequate supply ofblood to provide oxygen and nutrients to the brain tissue and to removewaste products from the brain tissue, brain cells rapidly begin to die.The symptoms of stroke are dependent on the region of the brain which isaffected by the inadequate blood flow. Symptoms include paralysis,numbness or weakness of the muscles, loss of balance, dizziness, suddensevere headaches, speech impairment, loss of memory, loss of reasoningability, sudden confusion, vision impairment, coma or even death. Astroke is also referred to as a brain attack or a cerebrovascularaccident (CVA). The symptoms of stroke may be brief if adequate bloodflow is restored within a short period of time. However, if inadequateblood flow continues for a significant period of time, the symptoms canbe permanent.

In some embodiments, the stroke is cerebral ischemia. Cerebral ischemiaresults when there is insufficient blood flow to the tissues of thebrain to meet metabolic demand. In some embodiments, the cerebralischemia is focal cerebral ischemia, i.e. confined to a specific regionof the brain. In some embodiments the cerebral ischemia is globalcerebral ischemia, i.e. encompassing a wide area of the brain tissue.Focal cerebral ischemia commonly occurs when a cerebral vessel hasbecome blocked, either partially or completely, reducing the flow ofblood to a specific region of the brain. In some embodiments the focalcerebral ischemia is ischemic stroke. In some embodiments, the ischemicstroke is thrombotic, i.e. caused by a thrombus or blood clot, whichdevelops in a cerebral vessel and restricts or blocks blood flow. Insome embodiments the ischemic stroke is a thrombotic stroke. In someembodiments, the ischemic stroke is embolic, i.e. caused by an embolus,or an unattached mass that travels through the bloodstream and restrictsor blocks blood flow at a site distant from its point of origin. In someembodiments the ischemic stroke is an embolic stroke. Global cerebralischemia commonly occurs when blood flow to the brain as a whole isblocked or reduced. In some embodiments the global cerebral ischemia iscaused by hypoperfusion, i.e. due to shock. In some embodiments theglobal cerebral ischemia is a result of a cardiac arrest.

In some embodiments the subject diagnosed with brain injury has sufferedcerebral ischemia. In some embodiments, the subject diagnosed with braininjury has suffered focal cerebral ischemia. In some embodiments, thesubject diagnosed with brain injury has suffered an ischemic stroke. Insome embodiments, the subject diagnosed with brain injury has suffered athrombotic stroke. In some embodiments, the subject diagnosed with braininjury has suffered an embolic stroke. In some embodiments, the subjectdiagnosed with brain injury has suffered global cerebral ischemia. Insome embodiments, the subject diagnosed with brain injury has sufferedhypoperfusion. In some embodiments, the subject diagnosed with braininjury has suffered a cardiac arrest.

In some embodiments, the compositions of the invention are for use intreating cerebral ischemia. In some embodiments, the compositions of theinvention are for use in treating focal cerebral ischemia. In someembodiments, the compositions of the invention are for use treatingischemic stroke. In some embodiments, the compositions of the inventionare for use in treating thrombotic stroke. In some embodiments, thecompositions of the invention are for use in treating embolic stroke. Insome embodiments, the compositions of the invention are for use intreating global cerebral ischemia. In some embodiments, the compositionsof the invention are for use in treating hypoperfusion.

In some embodiments, the stroke is hemorrhagic stroke. Hemorrhagicstroke is caused by bleeding into or around the brain resulting inswelling, pressure and damage to the cells and tissues of the brain.Hemorrhagic stroke is commonly a result of a weakened blood vessel thatruptures and bleeds into the surrounding brain. In some embodiments, thehemorrhagic stroke is an intracerebral hemorrhage, i.e. caused bybleeding within the brain tissue itself. In some embodiments theintracerebral hemorrhage is caused by an intraparenchymal hemorrhage. Insome embodiments the intracerebral hemorrhage is caused by anintraventricular hemorrhage. In some embodiments the hemorrhagic strokeis a subarachnoid hemorrhage i.e. bleeding that occurs outside of thebrain tissue but still within the skull. In some embodiments, thehemorrhagic stroke is a result of cerebral amyloid angiopathy. In someembodiments, the hemorrhagic stroke is a result of a brain aneurysm. Insome embodiments, the hemorrhagic stroke is a result of cerebralarteriovenous malformation (AVM).

In some embodiments the subject diagnosed with brain injury has sufferedhemorrhagic stroke. In some embodiments, the subject diagnosed withbrain injury has suffered an intracerebral hemorrhage. In someembodiments, the subject diagnosed with brain injury has suffered anintraparenchymal hemorrhage. In some embodiments, the subject diagnosedwith brain injury has suffered an intraventricular hemorrhage. In someembodiments, the subject diagnosed with brain injury has suffered asubarachnoid hemorrhage. In some embodiments, the subject diagnosed withbrain injury has suffered cerebral amyloid angiopathy. In someembodiments, the subject diagnosed with brain injury has suffered abrain aneurysm. In some embodiments, the subject diagnosed with braininjury has suffered cerebral AVM.

In some embodiments, the compositions of the invention are for use intreating hemorrhagic stroke. In some embodiments, the compositions ofthe invention are for use in treating an intracerebral hemorrhage. Insome embodiments, the compositions of the invention are for use intreating an intraparenchymal hemorrhage. In some embodiments, thecompositions of the invention are for use in treating anintraventricular hemorrhage. In some embodiments, the compositions ofthe invention are for use in treating a subarachnoid hemorrhage. In someembodiments, the compositions of the invention are for use in treating acerebral amyloid angiopathy. In some embodiments, the compositions ofthe invention are for use in treating a brain aneurysm. In someembodiments, the compositions of the invention are for use in treatingcerebral AVM.

Restoration of adequate blood flow to the brain after a period ofinterruption, though effective in alleviating the symptoms associatedwith stroke, can paradoxically result in further damage to the braintissue. During the period of interruption, the affected tissue suffersfrom a lack of oxygen and nutrients, and the sudden restoration of bloodflow can result in inflammation and oxidative damage through theinduction of oxidative stress. This is known as reperfusion injury, andis well documented not only following stroke, but also following a heartattack or other tissue damage when blood supply returns to the tissueafter a period of ischemia or lack of oxygen. In some embodiments thesubject diagnosed with brain injury has suffered from reperfusion injuryas a result of stroke. In some embodiments, the compositions of theinvention are for use in treating reperfusion injury as a result ofstroke.

A transient ischemic attack (TIA), often referred to as a mini-stroke,is a recognised warning sign for a more serious stroke. Subjects whohave suffered one or more TIAs are therefore at greater risk of stroke.In some embodiments the subject diagnosed with brain injury has suffereda TIA. In some embodiments, the compositions of the invention are foruse in treating a TIA. In some embodiments, the compositions of theinvention are for use in treating brain injury in a subject who hassuffered a TIA.

High blood pressure, high blood cholesterol, a familial history ofstroke, heart disease, diabetes, brain aneurysms, arteriovenousmalformations, sickle cell disease, vasculitis, bleeding disorders, useof nonsteroidal anti-inflammatory drugs (NSAIDs), smoking tobacco,drinking large amounts of alcohol, illegal drug use, obesity, lack ofphysical activity and an unhealthy diet are all considered to be riskfactors for stroke. In particular, lowering blood pressure has beenconclusively shown to prevent both ischemic and hemorrhagic strokes [37,38]. In some embodiments, the compositions of the invention are for usein treating brain injury in a subject who has at least one risk factorfor stroke. In some embodiments the subject has two risk factors forstroke. In some embodiments the subject has three risk factors forstroke. In some embodiments the subject has four risk factors forstroke. In some embodiments the subject has more than four risk factorsfor stroke. In some embodiments the subject has high blood pressure. Insome embodiments the subject has high blood cholesterol. In someembodiments the subject has a familial history of stroke. In someembodiments the subject has heart disease. In some embodiments thesubject has diabetes. In some embodiments the subject has a brainaneurysm. In some embodiments the subject has arteriovenousmalformations. In some embodiments the subject has vasculitis. In someembodiments the subject has sickle cell disease. In some embodiments thesubject has a bleeding disorder. In some embodiments the subject has ahistory of use of nonsteroidal anti-inflammatory drugs (NSAIDs). In someembodiments the subject smokes tobacco. In some embodiments the subjectdrinks large amounts of alcohol. In some embodiments the subject usesillegal drugs. In some embodiments the subject is obese. In someembodiments the subject is overweight. In some embodiments the subjecthas a lack of physical activity. In some embodiments the subject has anunhealthy diet.

The examples indicate that the compositions of the invention may beuseful for treating brain injury and aiding recovery when administeredbefore the injury event occurs. Therefore, the compositions of theinvention may be particularly useful for treating brain injury whenadministered to subjects at risk of brain injury, such as stroke.

In certain embodiments, the compositions of the invention are for use inreducing the damage caused by a potential brain injury, preferably astroke. The compositions may reduce the damage caused when they areadministered before the potential brain injury occurs, in particularwhen administered to a patient identified as at risk of a brain injury.

The examples indicate that the compositions of the invention may beuseful for treating brain injury and aiding recovery when administeredafter the injury event occurs. Therefore, the compositions of theinvention may be particularly useful for treating brain injury whenadministered to subjects following a brain injury, such as stroke.

In some embodiments, the compositions of the invention treat braininjury by reducing motoric damage. In some embodiments, the compositionsof the invention treat brain injury by improving motor function. In someembodiments, the compositions of the invention treat brain injury byimproving muscle strength. In some embodiments, the compositions of theinvention treat brain injury by improving memory. In some embodiments,the compositions of the invention treat brain injury by improving socialrecognition. In some embodiments, the compositions of the inventiontreat brain injury by improving neurological function.

Treatment of brain injury may refer to, for example, an alleviation ofthe severity of symptoms. Treatment of brain injury may also refer toreducing the neurological impairments following stroke. Compositions ofthe invention for use in treating stroke may be provided to the subjectin advance of the onset of stroke, for example in a patient identifiedas being at risk of stroke. Compositions of the invention for use intreating stroke may be provided after a stroke has occurred, forexample, during recovery. Compositions of the invention for use intreating stroke may be provided during the acute phase of recovery (i.e.up to one week after stroke). Compositions of the invention for use intreating stroke may be provided during the subacute phase of recovery(i.e. from one week up to three months after stroke). Compositions ofthe invention for use in treating stroke may be provided during thechronic phase of recovery (from three months after stroke).

In certain embodiments, the compositions of the invention are for use incombination with a secondary active agent. In certain embodiments, thecompositions of the invention are for use in combination with aspirin ortissue plasminogen activator (tPA). Other secondary agents include otherantiplatelets (such as clopidogrel), anticoagulants (such as heparins,warfarin, apixaban, dabigatran, edoxaban or rivaroxaban),antihypertensives (such as diuretics, ACE inhibitors, calcium channelblockers, beta-blockers or alpha-blockers) or statins. The compositionsof the invention may improve the patient's response to the secondaryactive agent.

In certain embodiments, the compositions of the invention reduce theeffect of ischemia on tissues. In certain embodiments, the compositionsof the invention reduce the amount of damage to tissues caused byischemia. In certain embodiments, the tissues damaged by ischemia arethe cerebral tissues. In certain embodiments, the compositions of theinvention reduce necrosis or the number of necrotic cells. In certainembodiments, the compositions of the invention reduce apoptosis or thenumber of apoptotic cells. In certain embodiments, the compositions ofthe invention reduce the number of necrotic and apoptotic cells. Incertain embodiments, the compositions of the invention prevent celldeath by necrosis and/or apoptosis. In certain embodiments, thecompositions of the invention prevent cell death by necrosis and/orapoptosis caused by ischemia. In certain embodiments, the compositionsof the invention improve the recovery of the tissue damaged by ischemia.In certain embodiments, the compositions of the invention improve thespeed of clearance of necrotic cells and/or apoptotic cells. In certainembodiments, the compositions of the invention improve the efficacy ofthe clearance of necrotic cells and/or apoptotic cells. In certainembodiments, the compositions of the invention improve the replacementand/or regeneration of cells within tissues. In certain embodiments, thecompositions of the invention improve the replacement and/orregeneration of cells within tissues damaged by ischemia. In certainembodiments, the compositions of the invention improve the overallhistology of the tissue (for example upon a biopsy).

Modes of Administration

Preferably, the compositions of the invention are to be administered tothe gastrointestinal tract in order to enable delivery to and/or partialor total colonisation of the intestine with the bacterial strain of theinvention. Generally, the compositions of the invention are administeredorally, but they may be administered rectally, intranasally, or viabuccal or sublingual routes.

In certain embodiments, the compositions of the invention may beadministered as a foam, as a spray or a gel.

In certain embodiments, the compositions of the invention may beadministered as a suppository, such as a rectal suppository, for examplein the form of a theobroma oil (cocoa butter), synthetic hard fat (e.g.suppocire, witepsol), glycero-gelatin, polyethylene glycol, or soapglycerin composition.

In certain embodiments, the composition of the invention is administeredto the gastrointestinal tract via a tube, such as a nasogastric tube,orogastric tube, gastric tube, jejunostomy tube (J tube), percutaneousendoscopic gastrostomy (PEG), or a port, such as a chest wall port thatprovides access to the stomach, jejunum and other suitable access ports.

The compositions of the invention may be administered once, or they maybe administered sequentially as part of a treatment regimen. In certainembodiments, the compositions of the invention are to be administereddaily.

In certain embodiments of the invention, treatment according to theinvention is accompanied by assessment of the patient's gut microbiota.Treatment may be repeated if delivery of and/or partial or totalcolonisation with the strain of the invention is not achieved such thatefficacy is not observed, or treatment may be ceased if delivery and/orpartial or total colonisation is successful and efficacy is observed.

In certain embodiments, the composition of the invention may beadministered to a pregnant animal, for example a mammal such as a humanin order to prevent an inflammatory or autoimmune disease developing inher child in utero and/or after it is born.

The compositions of the invention may be administered to a patient thathas been diagnosed with a neurodegenerative disease, or that has beenidentified as being at risk of a neurodegenerative disease.

The compositions may also be administered as a prophylactic measure toprevent the development of neurodegenerative disease in a healthypatient.

The compositions of the invention may be administered to a patient thathas been identified as having an abnormal gut microbiota. For example,the patient may have reduced or absent colonisation by Roseburia, and inparticular Roseburia hominis.

The compositions of the invention may be administered as a food product,such as a nutritional supplement.

Generally, the compositions of the invention are for the treatment ofhumans, although they may be used to treat animals including monogastricmammals such as poultry, pigs, cats, dogs, horses or rabbits. Thecompositions of the invention may be useful for enhancing the growth andperformance of animals. If administered to animals, oral gavage may beused.

Compositions

Generally, the composition of the invention comprises bacteria. Inpreferred embodiments of the invention, the composition is formulated infreeze-dried form. For example, the composition of the invention maycomprise granules or gelatin capsules, for example hard gelatincapsules, comprising a bacterial strain of the invention.

Preferably, the composition of the invention comprises lyophilisedbacteria. Lyophilisation of bacteria is a well-established procedure andrelevant guidance is available in, for example, references [39-41].

Alternatively, the composition of the invention may comprise a live,active bacterial culture.

In some embodiments, the bacterial strain in the composition of theinvention has not been inactivated, for example, has not beenheat-inactivated. In some embodiments, the bacterial strain in thecomposition of the invention has not been killed, for example, has notbeen heat-killed. In some embodiments, the bacterial strain in thecomposition of the invention has not been attenuated, for example, hasnot been heat-attenuated. For example, in some embodiments, thebacterial strain in the composition of the invention has not beenkilled, inactivated and/or attenuated. For example, in some embodiments,the bacterial strain in the composition of the invention is live. Forexample, in some embodiments, the bacterial strain in the composition ofthe invention is viable. For example, in some embodiments, the bacterialstrain in the composition of the invention is capable of partially ortotally colonising the intestine. For example, in some embodiments, thebacterial strain in the composition of the invention is viable andcapable of partially or totally colonising the intestine.

In some embodiments, the composition comprises a mixture of livebacterial strains and bacterial strains that have been killed.

In preferred embodiments, the composition of the invention isencapsulated to enable delivery of the bacterial strain to theintestine. Encapsulation protects the composition from degradation untildelivery at the target location through, for example, rupturing withchemical or physical stimuli such as pressure, enzymatic activity, orphysical disintegration, which may be triggered by changes in pH. Anyappropriate encapsulation method may be used. Exemplary encapsulationtechniques include entrapment within a porous matrix, attachment oradsorption on solid carrier surfaces, self-aggregation by flocculationor with cross-linking agents, and mechanical containment behind amicroporous membrane or a microcapsule. Guidance on encapsulation thatmay be useful for preparing compositions of the invention is availablein, for example, references [42] and [43].

The composition may be administered orally and may be in the form of atablet, capsule or powder. Encapsulated products are preferred becauseRoseburia are anaerobes. Other ingredients (such as vitamin C, forexample), may be included as oxygen scavengers and prebiotic substratesto improve the delivery and/or partial or total colonisation andsurvival in vivo. Alternatively, the probiotic composition of theinvention may be administered orally as a food or nutritional product,such as milk or whey based fermented dairy product, or as apharmaceutical product.

The composition may be formulated as a probiotic.

A composition of the invention includes a therapeutically effectiveamount of a bacterial strain of the invention. A therapeuticallyeffective amount of a bacterial strain is sufficient to exert abeneficial effect upon a patient. A therapeutically effective amount ofa bacterial strain may be sufficient to result in delivery to and/orpartial or total colonisation of the patient's intestine.

A suitable daily dose of the bacteria, for example for an adult human,may be from about 1×10³ to about 1×10¹¹ colony forming units (CFU); forexample, from about 1×10⁷ to about 1×10¹⁰ CFU; in another example fromabout 1×10⁶ to about 1×10¹⁰ CFU.

In certain embodiments, the composition contains the bacterial strain inan amount of from about 1×10⁶ to about 1×10¹¹ CFU/g, respect to theweight of the composition; for example, from about 1×10⁸ to about 1×10¹⁰CFU/g. The dose may be, for example, 1 g, 3 g, 5 g, and 10 g.

Typically, a probiotic, such as the composition of the invention, isoptionally combined with at least one suitable prebiotic compound. Aprebiotic compound is usually a non-digestible carbohydrate such as anoligo- or polysaccharide, or a sugar alcohol, which is not degraded orabsorbed in the upper digestive tract. Known prebiotics includecommercial products such as inulin and transgalacto-oligosaccharides.

In certain embodiments, the probiotic composition of the presentinvention includes a prebiotic compound in an amount of from about 1 toabout 30% by weight, respect to the total weight composition, (e.g. from5 to 20% by weight). Carbohydrates may be selected from the groupconsisting of: fructo-oligosaccharides (or FOS), short-chainfructo-oligosaccharides, inulin, isomalt-oligosaccharides, pectins,xylo-oligosaccharides (or XOS), chitosan-oligosaccharides (or COS),beta-glucans, arable gum modified and resistant starches, polydextrose,D-tagatose, acacia fibers, carob, oats, and citrus fibers. In oneaspect, the prebiotics are the short-chain fructo-oligosaccharides (forsimplicity shown herein below as FOSs-c.c); said FOSs-c.c. are notdigestible carbohydrates, generally obtained by the conversion of thebeet sugar and including a saccharose molecule to which three glucosemolecules are bonded.

In certain embodiments, the compositions of the invention are used incombination with another therapeutic compound for treating or preventingthe neurodegenerative disorder. In some embodiments, the compositions ofthe invention are administered with nutritional supplements thatmodulate neuroprotection or neuroproliferation. In preferredembodiments, the nutritional supplements comprise or consist ofnutritional vitamins. In certain embodiments, the vitamins are vitaminB6, magnesium, dimethylglycine (vitamin B16) and vitamin C. In certainembodiments, the compositions of the invention are administered incombination with another probiotic.

In certain embodiments, the compositions of the invention are for use inenhancing the effect of a second agent on a neurodegenerative disease.The immune modulatory effects of the compositions of the invention maymake the brain more susceptible to conventional therapies such asLevodopa, dopamine agonists, MAO-B inhibitors, COMT inhibitors,Glutamate antagonists, or anticholinergics, which are exemplarysecondary agents to be administered in combination (sequentially orcontemporaneously) with the compositions of the invention.

The compositions of the invention may comprise pharmaceuticallyacceptable excipients or carriers. Examples of such suitable excipientsmay be found in the reference [44]. Acceptable carriers or diluents fortherapeutic use are well known in the pharmaceutical art and aredescribed, for example, in reference [45]. Examples of suitable carriersinclude lactose, starch, glucose, methyl cellulose, magnesium stearate,mannitol, sorbitol and the like. Examples of suitable diluents includeethanol, glycerol and water. The choice of pharmaceutical carrier,excipient or diluent can be selected with regard to the intended routeof administration and standard pharmaceutical practice. Thepharmaceutical compositions may comprise as, or in addition to, thecarrier, excipient or diluent any suitable binder(s), lubricant(s),suspending agent(s), coating agent(s), solubilising agent(s). Examplesof suitable binders include starch, gelatin, natural sugars such asglucose, anhydrous lactose, free-flow lactose, beta-lactose, cornsweeteners, natural and synthetic gums, such as acacia, tragacanth orsodium alginate, carboxymethyl cellulose and polyethylene glycol.Examples of suitable lubricants include sodium oleate, sodium stearate,magnesium stearate, sodium benzoate, sodium acetate, sodium chloride andthe like. Preservatives, stabilizers, dyes and even flavouring agentsmay be provided in the pharmaceutical composition. Examples ofpreservatives include sodium benzoate, sorbic acid and esters ofp-hydroxybenzoic acid. Antioxidants and suspending agents may be alsoused.

The compositions of the invention may be formulated as a food product.For example, a food product may provide nutritional benefit in additionto the therapeutic effect of the invention, such as in a nutritionalsupplement. Similarly, a food product may be formulated to enhance thetaste of the composition of the invention or to make the compositionmore attractive to consume by being more similar to a common food item,rather than to a pharmaceutical composition. In certain embodiments, thecomposition of the invention is formulated as a milk-based product. Theterm “milk-based product” means any liquid or semi-solid milk- orwhey-based product having a varying fat content. The milk-based productcan be, e.g., cow's milk, goat's milk, sheep's milk, skimmed milk, wholemilk, milk recombined from powdered milk and whey without anyprocessing, or a processed product, such as yoghurt, curdled milk, curd,sour milk, sour whole milk, butter milk and other sour milk products.Another important group includes milk beverages, such as whey beverages,fermented milks, condensed milks, infant or baby milks; flavoured milks,ice cream; milk-containing food such as sweets.

In some embodiments, the compositions of the invention comprise one ormore bacterial strains of the genus Roseburia and do not containbacteria from any other genera, or which comprise only de minimis orbiologically irrelevant amounts of bacteria from another genera. Thus,in some embodiments, the invention provides a composition comprising oneor more bacterial strains of the genus Roseburia, which does not containbacteria from any other genera or which comprises only de minimis orbiologically irrelevant amounts of bacteria from another genera, for usein therapy.

In some embodiments, the compositions of the invention comprise one ormore bacterial strains of the species Roseburia hominis and do notcontain bacteria from any other species, or which comprise only deminimis or biologically irrelevant amounts of bacteria from anotherspecies. Thus, in some embodiments, the invention provides a compositioncomprising one or more bacterial strains of the species Roseburiahominis, which does not contain bacteria from any other species or whichcomprises only de minimis or biologically irrelevant amounts of bacteriafrom another species, for use in therapy.

In some embodiments, the compositions of the invention comprise one ormore bacterial strains of the species Roseburia intestinalis and do notcontain bacteria from any other species, or which comprise only deminimis or biologically irrelevant amounts of bacteria from anotherspecies. Thus, in some embodiments, the invention provides a compositioncomprising one or more bacterial strains of the species Roseburiaintestinalis, which does not contain bacteria from any other species orwhich comprises only de minimis or biologically irrelevant amounts ofbacteria from another species, for use in therapy.

In some embodiments, the compositions of the invention comprise one ormore bacterial strains of the species Roseburia faecis and do notcontain bacteria from any other species, or which comprise only deminimis or biologically irrelevant amounts of bacteria from anotherspecies. Thus, in some embodiments, the invention provides a compositioncomprising one or more bacterial strains of the species Roseburiafaecis, which does not contain bacteria from any other species or whichcomprises only de minimis or biologically irrelevant amounts of bacteriafrom another species, for use in therapy.

In some embodiments, the compositions of the invention comprise one ormore bacterial strains of the species Roseburia hominis and do notcontain bacteria from any other Roseburia species, or which compriseonly de minimis or biologically irrelevant amounts of bacteria fromanother Roseburia species. Thus, in some embodiments, the inventionprovides a composition comprising one or more bacterial strains of thespecies Roseburia hominis, which does not contain bacteria from anyother Roseburia species or which comprises only de minimis orbiologically irrelevant amounts of bacteria from another Roseburiaspecies, for use in therapy.

In some embodiments, the compositions of the invention comprise one ormore bacterial strains of the species Roseburia intestinalis and do notcontain bacteria from any other Roseburia species, or which compriseonly de minimis or biologically irrelevant amounts of bacteria fromanother Roseburia species. Thus, in some embodiments, the inventionprovides a composition comprising one or more bacterial strains of thespecies Roseburia intestinalis, which does not contain bacteria from anyother Roseburia species or which comprises only de minimis orbiologically irrelevant amounts of bacteria from another Roseburiaspecies, for use in therapy.

In some embodiments, the compositions of the invention comprise one ormore bacterial strains of the species Roseburia faecis and do notcontain bacteria from any other Roseburia species, or which compriseonly de minimis or biologically irrelevant amounts of bacteria fromanother Roseburia species. Thus, in some embodiments, the inventionprovides a composition comprising one or more bacterial strains of thespecies Roseburia faecis, which does not contain bacteria from any otherRoseburia species or which comprises only de minimis or biologicallyirrelevant amounts of bacteria from another Roseburia species, for usein therapy.

In certain embodiments, the compositions of the invention contain asingle bacterial strain or species and do not contain any otherbacterial strains or species. Such compositions may comprise only deminimis or biologically irrelevant amounts of other bacterial strains orspecies. Such compositions may be a culture that is substantially freefrom other species of organism.

In some embodiments, the invention provides a composition comprising asingle bacterial strain of the genus Roseburia, which does not containbacteria from any other strains or which comprises only de minimis orbiologically irrelevant amounts of bacteria from another strain for usein therapy.

In some embodiments, the invention provides a composition comprising asingle bacterial strain of the species Roseburia hominis, which does notcontain bacteria from any other strains or which comprises only deminimis or biologically irrelevant amounts of bacteria from anotherstrain for use in therapy.

In some embodiments, the invention provides a composition comprising asingle bacterial strain of the species Roseburia intestinalis, whichdoes not contain bacteria from any other strains or which comprises onlyde minimis or biologically irrelevant amounts of bacteria from anotherstrain for use in therapy.

In some embodiments, the invention provides a composition comprising asingle bacterial strain of the species Roseburia faecis, which does notcontain bacteria from any other strains or which comprises only deminimis or biologically irrelevant amounts of bacteria from anotherstrain for use in therapy.

In some embodiments, the compositions of the invention comprise morethan one bacterial strain. For example, in some embodiments, thecompositions of the invention comprise more than one strain from withinthe same species (e.g. more than 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20,25, 30, 35, 40 or 45 strains), and, optionally, do not contain bacteriafrom any other species. In some embodiments, the compositions of theinvention comprise less than 50 strains from within the same species(e.g. less than 45, 40, 35, 30, 25, 20, 15, 12, 10, 9, 8, 7, 6, 5, 4 or3 strains), and, optionally, do not contain bacteria from any otherspecies. In some embodiments, the compositions of the invention comprise1-40, 1-30, 1-20, 1-19, 1-18, 1-15, 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 1-4,1-3, 1-2, 2-50, 2-40, 2-30, 2-20, 2-15, 2-10, 2-5, 6-30, 6-15, 16-25, or31-50 strains from within the same species and, optionally, do notcontain bacteria from any other species. The invention comprises anycombination of the foregoing.

In some embodiments, the composition comprises a microbial consortium.For example, in some embodiments, the composition comprises theRoseburia bacterial strain as part of a microbial consortium. Forexample, in some embodiments, the Roseburia bacterial strain is presentin combination with one or more (e.g. at least 2, 3, 4, 5, 10, 15 or 20)other bacterial strains from other genera with which it can livesymbiotically in vivo in the intestine. For example, in someembodiments, the composition comprises a bacterial strain of Roseburiain combination with a bacterial strain from a different genus. In someembodiments, the microbial consortium comprises two or more bacterialstrains obtained from a faeces sample of a single organism, e.g. ahuman. In some embodiments, the microbial consortium is not foundtogether in nature. For example, in some embodiments, the microbialconsortium comprises bacterial strains obtained from faeces samples ofat least two different organisms. In some embodiments, the two differentorganisms are from the same species, e.g. two different humans. In someembodiments, the two different organisms are an infant human and anadult human. In some embodiments, the two different organisms are ahuman and a non-human mammal.

In some embodiments, the composition of the invention additionallycomprises a bacterial strain that has the same safety and therapeuticefficacy characteristics as strain MRx0001, but which is not MRx0001, orwhich is not a Roseburia hominis.

In some embodiments, the composition of the invention additionallycomprises a bacterial strain that has the same safety and therapeuticefficacy characteristics as strain A, but which is not strain A, orwhich is not a Roseburia intestinalis.

In some embodiments in which the composition of the invention comprisesmore than one bacterial strain, species or genus, the individualbacterial strains, species or genera may be for separate, simultaneousor sequential administration. For example, the composition may compriseall of the more than one bacterial strain, species or genera, or thebacterial strains, species or genera may be stored separately and beadministered separately, simultaneously or sequentially. In someembodiments, the more than one bacterial strains, species or genera arestored separately but are mixed together prior to use.

In some embodiments, the bacterial strain for use in the invention isobtained from human adult faeces. In some embodiments in which thecomposition of the invention comprises more than one bacterial strain,all of the bacterial strains are obtained from human adult faeces or ifother bacterial strains are present they are present only in de minimisamounts. The bacteria may have been cultured subsequent to beingobtained from the human adult faeces and being used in a composition ofthe invention.

In some embodiments, the bacterial strain for use in the invention isobtained from human infant faeces. In some embodiments in which thecomposition of the invention comprises more than one bacterial strain,all of the bacterial strains are obtained from human infant faeces or ifother bacterial strains are present they are present only in de minimisamounts. The bacteria may have been cultured subsequent to beingobtained from the human infant faeces and being used in a composition ofthe invention.

As mentioned above, in some embodiments, the one or more Roseburiabacterial strains is/are the only therapeutically active agent(s) in acomposition of the invention. In some embodiments, the bacterialstrain(s) in the composition is/are the only therapeutically activeagent(s) in a composition of the invention.

The compositions for use in accordance with the invention may or may notrequire marketing approval.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, wherein said bacterial strain is lyophilised. In certainembodiments, the invention provides the above pharmaceuticalcomposition, wherein said bacterial strain is spray dried. In certainembodiments, the invention provides the above pharmaceuticalcomposition, wherein the bacterial strain is lyophilised or spray driedand wherein it is live. In certain embodiments, the invention providesthe above pharmaceutical composition, wherein the bacterial strain islyophilised or spray dried and wherein it is viable. In certainembodiments, the invention provides the above pharmaceuticalcomposition, wherein the bacterial strain is lyophilised or spray driedand wherein it is capable of partially or totally colonising theintestine. In certain embodiments, the invention provides the abovepharmaceutical composition, wherein the bacterial strain is lyophilisedor spray dried and wherein it is viable and capable of partially ortotally colonising the intestine.

In some cases, the lyophilised bacterial strain is reconstituted priorto administration. In some cases, the reconstitution is by use of adiluent described herein.

The compositions of the invention can comprise pharmaceuticallyacceptable excipients, diluents or carriers.

In certain embodiments, the invention provides a pharmaceuticalcomposition comprising: a bacterial strain of the invention; and apharmaceutically acceptable excipient, carrier or diluent; wherein thebacterial strain is in an amount sufficient to treat a neurodegenerativedisorder when administered to a subject in need thereof.

In certain embodiments, the invention provides pharmaceuticalcomposition comprising: a bacterial strain of the invention; and apharmaceutically acceptable excipient, carrier or diluent; wherein thebacterial strain is in an amount sufficient to treat or prevent aneurodegenerative disorder.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, wherein the amount of the bacterial strain is from about1×10³ to about 1×10¹¹ colony forming units per gram with respect to aweight of the composition.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, wherein the composition is administered at a dose of 1 g, 3g, 5 g or 10 g.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, wherein the composition is administered by a methodselected from the group consisting of oral, rectal, subcutaneous, nasal,buccal, and sublingual.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, comprising a carrier selected from the group consisting oflactose, starch, glucose, methyl cellulose, magnesium stearate, mannitoland sorbitol.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, comprising a diluent selected from the group consisting ofethanol, glycerol and water.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, comprising an excipient selected from the group consistingof starch, gelatin, glucose, anhydrous lactose, free-flow lactose,beta-lactose, corn sweetener, acacia, tragacanth, sodium alginate,carboxymethyl cellulose, polyethylene glycol, sodium oleate, sodiumstearate, magnesium stearate, sodium benzoate, sodium acetate and sodiumchloride.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, further comprising at least one of a preservative, anantioxidant and a stabilizer.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, comprising a preservative selected from the groupconsisting of sodium benzoate, sorbic acid and esters ofp-hydroxybenzoic acid.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, wherein said bacterial strain is lyophilised.

In certain embodiments, the invention provides the above pharmaceuticalcomposition, wherein when the composition is stored in a sealedcontainer at about 4.0 or about 25.0 and the container is placed in anatmosphere having 50% relative humidity, at least 80% of the bacterialstrain as measured in colony forming units, remains after a period of atleast about: 1 month, 3 months, 6 months, 1 year, 1.5 years, 2 years,2.5 years or 3 years.

In some embodiments, the composition of the invention is provided in asealed container comprising a composition as described herein. In someembodiments, the sealed container is a sachet or bottle. In someembodiments, the composition of the invention is provided in a syringecomprising a composition as described herein.

The composition of the present invention may, in some embodiments, beprovided as a pharmaceutical formulation. For example, the compositionmay be provided as a tablet or capsule. In some embodiments, the capsuleis a gelatine capsule (“gel-cap”).

In some embodiments, the compositions of the invention are administeredorally. Oral administration may involve swallowing, so that the compoundenters the gastrointestinal tract, and/or buccal, lingual, or sublingualadministration by which the compound enters the blood stream directlyfrom the mouth.

Pharmaceutical formulations suitable for oral administration includesolid plugs, solid microparticulates, semi-solid and liquid (includingmultiple phases or dispersed systems) such as tablets; soft or hardcapsules containing multi- or nano-particulates, liquids (e.g. aqueoussolutions), emulsions or powders; lozenges (including liquid-filled);chews; gels; fast dispersing dosage forms; films; ovules; sprays; andbuccal/mucoadhesive patches.

In some embodiments the pharmaceutical formulation is an entericformulation, i.e. a gastro-resistant formulation (for example, resistantto gastric pH) that is suitable for delivery of the composition of theinvention to the intestine by oral administration. Enteric formulationsmay be particularly useful when the bacteria or another component of thecomposition is acid-sensitive, e.g. prone to degradation under gastricconditions.

In some embodiments, the enteric formulation comprises an entericcoating. In some embodiments, the formulation is an enteric-coateddosage form. For example, the formulation may be an enteric-coatedtablet or an enteric-coated capsule, or the like. The enteric coatingmay be a conventional enteric coating, for example, a conventionalcoating for a tablet, capsule, or the like for oral delivery. Theformulation may comprise a film coating, for example, a thin film layerof an enteric polymer, e.g. an acid-insoluble polymer.

In some embodiments, the enteric formulation is intrinsically enteric,for example, gastro-resistant without the need for an enteric coating.Thus, in some embodiments, the formulation is an enteric formulationthat does not comprise an enteric coating. In some embodiments, theformulation is a capsule made from a thermogelling material. In someembodiments, the thermogelling material is a cellulosic material, suchas methylcellulose, hydroxymethylcellulose orhydroxypropylmethylcellulose (HPMC). In some embodiments, the capsulecomprises a shell that does not contain any film forming polymer. Insome embodiments, the capsule comprises a shell and the shell compriseshydroxypropylmethylcellulose and does not comprise any film formingpolymer (e.g. see [46]). In some embodiments, the formulation is anintrinsically enteric capsule (for example, Vcaps® from Capsugel).

In some embodiments, the formulation is a soft capsule. Soft capsulesare capsules which may, owing to additions of softeners, such as, forexample, glycerol, sorbitol, maltitol and polyethylene glycols, presentin the capsule shell, have a certain elasticity and softness. Softcapsules can be produced, for example, on the basis of gelatine orstarch. Gelatine-based soft capsules are commercially available fromvarious suppliers. Depending on the method of administration, such as,for example, orally or rectally, soft capsules can have various shapes,they can be, for example, round, oval, oblong or torpedo-shaped. Softcapsules can be produced by conventional processes, such as, forexample, by the Scherer process, the Accogel process or the droplet orblowing process.

Culturing Methods

The bacterial strains for use in the present invention can be culturedusing standard microbiology techniques as detailed in, for example,references [47-49].

The solid or liquid medium used for culture may be YCFA agar or YCFAmedium. YCFA medium may include (per 100 ml, approximate values):Casitone (1.0 g), yeast extract (0.25 g), NaHCO₃ (0.4 g), cysteine (0.1g), K₂HPO₄ (0.045 g), KH₂PO₄ (0.045 g), NaCl (0.09 g), (NH₄)₂SO₄ (0.09g), MgSO₄.7H₂O (0.009 g), CaCl₂) (0.009 g), resazurin (0.1 mg), hemin (1mg), biotin (1 μg), cobalamin (1 μg), p-aminobenzoic acid (3 μg), folicacid (5 μg), and pyridoxamine (15 μg).

Bacterial Strains for Use in Vaccine Compositions

The inventors have identified that the bacterial strains of theinvention are useful for treating or preventing neurodegenerativedisorders. This is likely to be a result of the effect that thebacterial strains of the invention have on the host immune system.Therefore, the compositions of the invention may also be useful forpreventing neurodegenerative disorders, when administered as vaccinecompositions. In certain such embodiments, the bacterial strains of theinvention may be killed, inactivated or attenuated. In certain suchembodiments, the compositions may comprise a vaccine adjuvant. Incertain embodiments, the compositions are for administration viainjection, such as via subcutaneous injection.

General

The practice of the present invention will employ, unless otherwiseindicated, conventional methods of chemistry, biochemistry, molecularbiology, immunology and pharmacology, within the skill of the art. Suchtechniques are explained fully in the literature. See, e.g., references[50] and [51-57], etc.

The term “comprising” encompasses “including” as well as “consisting”e.g. a composition “comprising” X may consist exclusively of X or mayinclude something additional e.g. X+Y.

The term “about” in relation to a numerical value x is optional andmeans, for example, x±10%.

The word “substantially” does not exclude “completely” e.g. acomposition which is “substantially free” from Y may be completely freefrom Y. Where necessary, the word “substantially” may be omitted fromthe definition of the invention.

References to a percentage sequence identity between two nucleotidesequences means that, when aligned, that percentage of nucleotides arethe same in comparing the two sequences. This alignment and the percenthomology or sequence identity can be determined using software programsknown in the art, for example those described in section 7.7.18 of ref[58]. A preferred alignment is determined by the Smith-Waterman homologysearch algorithm using an affine gap search with a gap open penalty of12 and a gap extension penalty of 2, BLOSUM matrix of 62. TheSmith-Waterman homology search algorithm is disclosed in ref. [59].

Unless specifically stated, a process or method comprising numeroussteps may comprise additional steps at the beginning or end of themethod, or may comprise additional intervening steps. Also, steps may becombined, omitted or performed in an alternative order, if appropriate.

Various embodiments of the invention are described herein. It will beappreciated that the features specified in each embodiment may becombined with other specified features, to provide further embodiments.In particular, embodiments highlighted herein as being suitable, typicalor preferred may be combined with each other (except when they aremutually exclusive).

MODES FOR CARRYING OUT THE INVENTION Example 1—Efficacy of BacterialInocula to Reduce IL-6 Secretion Summary

Activation of proinflammatory cytokines has been associated with neurondamage in neurodegenerative disease. Lipopolysaccharide (LPS) is a knownstimulator of the proinflammatory cytokine IL-6. Human glioblastomaastrocytoma cells were treated with compositions comprising bacterialstrains according to the invention in combination with LPS to observetheir ability to modulate the levels of IL-6.

Material and Methods Bacterial Strain

Roseburia hominis MRx0001

Cell Line

MG U373 is a human glioblastoma astrocytoma derived from a malignanttumour and were purchased from Sigma-Aldrich (cat n. 08061901-1VL). MGU373 human glioblastoma astrocytoma cells were grown in MEM (SigmaAldrich, cat n. M-2279) supplemented with 10% FBS, 1% Pen Strep, 4 mML-Glut, 1×MEM Non essential Amino Acid solution and 1× Sodium Piruvate.

Method

Once grown the MG U373 cells were plated on 24-well plate at 100,000cells/well. The cells were treated with LPS (1 ug/mL) alone or with 10%of bacteria supernatant from MRx0001 for 24h. A control was alsoperformed where the cells were incubated in untreated media. The cellswere treated with 10% of bacteria supernatant from MRx0001 or YCFA alonefor 24h. A control was also performed where the cells were incubated inuntreated media. Afterwards the cell free supernatants were collected,centrifuged at 10,000 g for 3 min at 4° C. IL-6 was measured using theHuman IL-6 ELISA Kit from Peprotech (cat n.#900-K16) according tomanufacturer instruction.

Results

The results of these experiments are shown in FIG. 1 and FIG. 9.Treatment of neuroblastoma cells with LPS and the bacteria strain led toa decrease in the level of IL-6 secreted to the same level as thecontrol cells that were untreated with LPS (FIG. 1). Treatment ofneuroblastoma cells with Roseburia hominis led to a decrease in thelevel of IL-6 secreted below the levels observed with the cells treatedwith the control (FIG. 9).

Example 2—Efficacy of Bacterial Inocula to Reduce NFκB ActivationSummary

Activation of the NFκB promoter leads to the production ofproinflammatory cytokines including IL-1β, IL-1α, IL-18, TNFα and IL-6.The NFκB promoter can be activated by α-synuclein and LPS by stimulatingthe TLR4 ligand. Mutations in α-synuclein, such as α-synuclein A53T, areimplicated in familial Parkinson's. Treatment of neuronal cells with LPSsimulates Parkinson's caused by environmental factors. The ability ofcompositions comprising bacterial strains according to the invention toinhibit the activation of the NFκB promoter was investigated.

Material and Methods Bacterial Strain

Roseburia hominis MRx0001

Cell Line

Human Hek blue TLR4 were purchased from InvivoGen (cat n. hkb-htlr4).Human Hek blue TLR4 were grown in DMEM high glucose (Sigma Aldrich, catn. D-6171) supplemented with 10% FBS, 1% Pen Strep, 4 mM L-Glut,Normocin and 1×FMK Blue selection solution.

Method

Once grown the Human Hek blue cells were plated in 96 well plate at25,000 cells/well in 4 replicates. One set of cells were treated withα-synuclein A53T (1 ug/mL) alone or with 10% of bacteria supernatantfrom MRx0001 for 22h. The second set of cells were treated with LPS (10ng/mL, from Salmonella enterica serotype Typhimurium, Sigma Aldrich, catn. L6143) alone or with 10% of bacteria supernatant from MRx0001 for22h. The cells were subsequently spun down and 20 ul of the supernatantwas mixed with 200 ul of Quanti Blue reagent (InvivoGen, cat n.rep-qb2), incubated for 2 h and absorbance read at 655 nm.

Results

The results of these experiments are shown in FIGS. 2 and 3. FIG. 2shows that the activation of the NFκB promoter by α-synuclein isinhibited by MRx0001. FIG. 3 shows that the activation of the NFκBpromoter by LPS is inhibited by MRx0001.

Example 3—Efficacy of Bacterial Inocula to Alter Antioxidant CapacitySummary

The ability of compositions comprising bacterial strains according tothe invention to alter the antioxidant capacity. The antioxidantcapacity of the bacterial strain was established using the well-knownABTS (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid)) assay.

Bacterial Strain

Roseburia hominis MRx0001

Method

Bacterial cells (10⁶ or greater) were collected and centrifuged. Theywere resuspended in assay buffer (using three times the pellet volume).The suspension was sonicated on ice for 5 minutes and then spun down at12,000×g for 10 minutes. The supernatant was removed and measured usingthe ABTS assay kit produced by Sigma Aldrich (code CS0790), inaccordance with manufacturer's instructions.

Results

The results of these experiments are shown in FIG. 4. FIG. 4 shows thatMRx0001 has an antioxidant capacity of approximately 1 mM compared toTrolox.

Example 4—Efficacy of Bacterial Inocula to Alter Lipid PeroxidationLevels Summary

The ability of compositions comprising bacterial strains according tothe invention to alter lipid peroxidation levels was investigated. Thethiobarbituric reactive substances assay (TBARs) was used to measure theby-products of lipid peroxidation.

Material and Methods Bacterial Strain

Roseburia hominis MRx0001

Method

Bacterial cells (10⁶ or greater) were collected and centrifuged, a washstep was performed with isotonic saline before the pellet wasre-suspended in potassium chloride assay buffer. The suspension wassonicated on ice for 10 minutes and then spun down at 10,000×g for 10minutes. The supernatant was removed and the level of lipid peroxidationevaluated using the thiobarbituric reactive substances assay.

Results

The results of the experiments are shown in FIG. 5. FIG. 5 shows thatMRx0001 is able to inhibit lipid peroxidation by approximately 18%,which is a higher antioxidant capacity than the positive control,butylated hydroxytoluene (1% w/v).

Example 5—Efficacy of Bacterial Inocula on Histone DeacetylataseActivity Summary

The ability of compositions comprising bacterial strains according tothe invention to alter histone deacetylatase activity was investigated.Dysregulation of histone deacetylatase has been implicated in thepathogenesis associated with age-associated neurodegenerative diseases.

Material and Methods Bacterial Strain

Roseburia hominis MRx0001

Cell Line

The cell line HT-29 was used because histone deacetylase is present.

Method

Cell free supernatants of stationary phase bacterial cultures wereisolated by centrifugation and filtering in a 0.22 uM filter. HT-29cells were used 3 days' post confluence and stepped down in 1 mL DTS 24hours prior to commencement of the experiment. The HT-29 cells werechallenged with 10% cell free supernatant diluted in DTS and this wasleft to incubate for 48 hours. Nuclease proteins were then extractedusing the Sigma Aldrich Nuclease extraction kit and samples were snapfrozen prior to HDAC activity measurement. HDAC activity was assessedfluorometrically using the Sigma Aldrich (UK) kit.

Results

The results of the experiments are shown in FIG. 6. FIG. 6 shows thatMRx0001 is able reduce the levels of histone deacetylase activity.

Example 6—Level of Indole Production in Bacteria Summary

The ability of the bacteria of the invention to produce indole wasinvestigated. Indole has been implicated in attenuating inflammation andoxidative stress.

Material and Methods Bacterial Strain

Roseburia hominis MRx0001

ATCC 11775 is a bacterial reference strain that is known to produceindole.

Method

Intact bacterial cells in stationary phase were incubated with 6 mMTryptophan for 48 hours. Bacterial species which possess the enzymetryptophanase will utilise tryptophan as a substrate to produce indole.Following the 48 hour incubation period, the supernatant was removed andadded to Kovac's reagent for quantification of indole. Standards, stocksolutions and reagents were prepared using standardised methodsvalidated in-house.

Results

The results of the experiments are shown in FIG. 7. FIG. 7 shows thatMRx0001 has the capacity to produce indole from tryptophan, atconcentrations of approximately 0.25 mM.

Example 7—Level of Kynurenine Production in Bacteria Summary

The ability of the bacteria of the invention to produce kynurenine wasinvestigated. Dysregulation of the kynurenine pathway can lead toactivation of the immune system and the accumulation of potentiallyneurotoxic compounds. Alterations in the kynurenine metabolism may beinvolved in the development of Parkinson's diseases.

Bacterial Strain

Roseburia hominis MRx0001

DSM 17136 is a strain of Bacteroides copricola that is known to producekynurenine.

Method

Cell free supernatants of stationary phase bacterial cultures wereisolated by centrifugation and filtering in a 0.22 uM filter and frozenuntil use. Kynurenine standards, stock solutions and reagents wereprepared using standardised methods validated in-house. Sample weretreated with trichloroacetic acid and centrifuged at 10,000×g for 10minutes at 4° C. The supernatant was collected and dispensed into a 96well plate. Ehrlich's reagent was used for kynurenine detection andadded at a ratio of 1:1.

Results

The results of the experiments are shown in FIG. 8. FIG. 8 shows thatMRx0001 has the capacity to produce kynurenine at a concentration ofapproximately 50 μM.

Example 8—Stability Testing

A composition described herein containing at least one bacterial straindescribed herein is stored in a sealed container at 25° C. or 4° C. andthe container is placed in an atmosphere having 30%, 40%, 50%, 60%, 70%,75%, 80%, 90% or 95% relative humidity. After 1 month, 2 months, 3months, 6 months, 1 year, 1.5 years, 2 years, 2.5 years or 3 years, atleast 50%, 60%, 70%, 80% or 90% of the bacterial strain shall remain asmeasured in colony forming units determined by standard protocols.

Example 9—Efficacy of Bacterial Inocula to Reduce IL-6 Secretion Summary

Activation of proinflammatory cytokines has been associated with neurondamage in neurodegenerative disease. Lipopolysaccharide (LPS) is a knownstimulator of the proinflammatory cytokine IL-6. Human glioblastomaastrocytoma cells were treated with compositions comprising bacterialstrains according to the invention in combination with LPS to observetheir ability to modulate the levels of IL-6.

Material and Methods Bacterial Strain

Roseburia intestinalis Strain ARoseburia faecis Strain B

Cell Line

MG U373 is a human glioblastoma astrocytoma derived from a malignanttumour and were purchased from Sigma-Aldrich (cat n. 08061901-1VL). MGU373 human glioblastoma astrocytoma cells were grown in MEM (SigmaAldrich, cat n. M-2279) supplemented with 10% FBS, 1% Pen Strep, 4 mML-Glut, 1×MEM Non essential Amino Acid solution and 1× Sodium Piruvate.

Method

Once grown the MG U373 cells were plated on 24-well plate at 100,000cells/well. The cells were treated with the following conditions for24h:

-   -   LPS (1 ug/mL)    -   LPS with 10% of bacteria supernatant from Strain A    -   LPS with 10% of bacteria supernatant from Strain B    -   LPS with YCFA media    -   YCFA

A control was also performed where the cells were incubated in untreatedmedia.

Afterwards the cell free supernatants were collected, centrifuged at10,000 g for 3 min at 4° C. IL-6 was measured using the Human IL-6 ELISAKit from Peprotech (cat n.#900-K16) according to manufacturerinstruction.

Results

The results of these experiments are shown in FIGS. 10 and 12. Treatmentof neuroblastoma cells with LPS and the bacteria strain A and B led to adecrease in the level of IL-6 secreted to the same level as the controlcells that were untreated with LPS (FIGS. 10A and 12A).

Treatment of neuroblastoma cells with Roseburia intestinalis andRoseburia faecis led to a decrease in the level of IL-6 secreted belowthe levels observed with the cells treated with the control (FIGS. 10Band 12B).

Example 10—Efficacy of Bacterial Inocula to Reduce NFκB ActivationSummary

Activation of the NFκB promoter leads to the production ofproinflammatory cytokines including IL-1β, IL-1α, IL-18, TNFα and IL-6.The NFκB promoter can be activated by α-synuclein and LPS by stimulatingthe TLR4 ligand. Mutations in α-synuclein, such as α-synuclein A53T, areimplicated in familial Parkinson's. Treatment of neuronal cells with LPSsimulates Parkinson's caused by environmental factors. The ability ofcompositions comprising bacterial strains according to the invention toinhibit the activation of the NFκB promoter was investigated.

Material and Methods Bacterial Strain

Roseburia intestinalis Strain ARoseburia faecis Strain B

Cell Line

Human Hek blue TLR4 were purchased from InvivoGen (cat n. hkb-hdr4).Human Hek blue TLR4 were grown in DMEM high glucose (Sigma Aldrich, catn. D-6171) supplemented with 10% FBS, 1% Pen Strep, 4 mM L-Glut,Normocin and 1×HEK Blue selection solution.

Method

Once grown the Human Hek blue cells were plated in 96 well plate at25,000 cells/well in 4 replicates. One set of cells were treated withα-synuclein A53T (1 ug/mL) alone or with 10% of bacteria supernatantfrom Strain A or Strain B for 22h. The second set of cells were treatedwith LPS (10 ng/mL, from Salmonella enterica serotype Typhimurium, SigmaAldrich, cat n. L6143) alone or with 10% of bacteria supernatant fromStrain A or Strain B for 22h. The cells were subsequently spun down and20 ul of the supernatant was mixed with 200 ul of Quanti Blue reagent(InvivoGen, cat n. rep-qb2), incubated for 2 h and absorbance read at655 nm.

Results

The results of these experiments are shown in FIGS. 11 and 13.

Sequences(Roseburia hominis strain A2-181 16S ribosomal RNA gene, partialsequence - AY804148) SEQ ID NO: 1   1 taaaggttga tcctggctca ggatgaacgc tggaggcgtg cttaacacat gcaagtcgaa  61 cgaagcactt taattgattt cttcggaatg aagtttttgt gactgagtgg cggacgggtg 121 agtaacgcgt gggtaacctc gctcatacag ggggataaca gttggaaacg actgctaata 181 ccgcataagc gcacaggatt gcatgatcca gtgtgaaaaa ctccggtggt atgagatgga 241 cccgcgtctg attagccagt tggcggggta acggcccacc aaagcgacga tcagtagccg 301 acctgagagg gtgaccggcc acattgggac tgagacacgg cccaaactcc tacgggaggc 361 agcagtgggt aatattgcac aatgggggaa accctgatgc agcgacgccg agtgagcgaa 421 gaagtatttc ggtatgtaaa gctctatcag caggaagaag aatgacggta cctgactaaa 481 aagcaccggc taaatacgtg ccagcagccg cggtaatacg tatggtgcaa gcgttatccg 541 gatttactgg gtgtaaaggg agcgcaggcg gtacggcaag tctgatgtga aatcccgggg 601 ctcaaccccg gtactgcatt ggaaactgtc ggactagggt gtctgagggg taagtggaat 661 tcctagtgta gcggtgaaat gcgtagatat taggaggaac accagtggcg aaggcggctt 721 actggacgat tactgacgct gaggctcgaa agcgtgggga gcaaacagga ttagataccc 781 tggtagtcca cgccgtaaac gatgaatact aggtgtcggg gagcattgct cttcggtgcc 841 gcagcaaacg caataagtat tccacctggg gagtacgttc gcaagaatga aactcaaagg 901 aattgacggg gacccgcaca agcggtggag catgtggttt aattcgaagc aacgcgaaga 961 accttaccaa gtcttgacat cccactgaca aagtatgtaa tgtactttct cttcggagca1021 gtggtgacag gtggtgcatg gttgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc1081 cgcaacgagc gcaaccccta ttcttagtag ccagcggttt ggccgggcac tctagggaga1141 ctgccaggga taacctggag gaaggtgggg atgacgtcaa atcatcatgc cccttatgac1201 ttgggctaca cacgtgctac aatggcgtaa acaaagggaa gcaatcccgc gagggggagc1261 aaatctcaaa aataacgtct cagttcggac tgtagtctgc aactcgacta cacgaagctg1321 gaatcgctag taatcgcgaa tcagaatgtc gcggtgaata cgttcccggg tcttgtacac1381 accgcccgtc acaccatggg agttggtaat gcccgaagtc agtgacccaa ccgcaaggag1441 ggagctgccg aagcaggact gataactggg gtgaagtcgt aacaagt(Roseburia hominis A2-183 16S rRNA gene, type strain A2-183T - AJ270482)SEQ ID NO: 2   1 gatcctggct caggatgaac gctggcggcg tgcttaacac atgcaagtcg aacgaagcac  61 tttaattgat ttcttcggaa tgaagttttt gtgactgagt ggcggacggg tgagtaacgc 121 gtgggtaacc tgcctcatac agggggataa cagttggaaa cgactgctaa taccgcataa 181 gcgcacagga ttgcatgatc cagtgtgaaa aactccggtg gtatgagatg gacccgcgtc 241 tgattagcca gttggcgggg taacggccca ccaaagcgac gatcagtagc cgacctgaga 301 gggtgaccgg ccacattggg actgagacac ggcccaaact cctacgggag gcagcagtgg 361 ggaatattgc acaatggggg aaaccctgat gcagcgacgc cgcgtgagcg aagaagtatt 421 tcggtatgta aagctctatc agcagggaag aagaatgcgg tacctgacta agaagcaccg 481 gctaaatacg tgccagcagc cgcggtaata cgtatggtgc aagcgttatc cggatttact 541 gggtgtaaag ggagcgcagg cggtacggca agtctgatgt gaaatcccgg ggctcaaccc 601 cggtactgca ttggaaactg tcggactaga gtgtcggagg ggtaagtgga attcctagtg 661 tagcggtgaa atgcgtagat attaggagga acaccagtgg cgaaggcggc ttactggacg 721 attactgacg ctgaggctcg aaagcgtggg gagcaaacag gattagatac cctggtagtc 781 cacgccgtaa acgatgaata ctaggtgtcg gggagcattg ctcttcggtg ccgcagcaaa 841 cgcaataagt attccacctg gggagtacgt tcgcaagaat gaaactcaaa ggaattgacg 901 gggacccgca caagcggtgg agcatgtggt ttaattcgaa gcaacgcgaa gaaccttacc 961 aagtcttgac atcccactga cagagtatgt aatgtacttt ctcttcggag cagtggtgac1021 aggtggtgca tggttgtcgt cagctcgtgt cgtgagatgt tgggttaagt cccgcaacga1081 gcgcaacccc tattcttagt agccagcggt tcggccgggc actctaggga gactgccagg1141 gataacctgg aggaaggtgg ggatgacgtc aaatcatcat gccccttatg acttgggcta1201 cacacgtgct acaatggcgt aaacaaaggg aagcaatccc gcgaggggga gcaaatctca1261 aaaataacgt ctcagttcgg actgtagtct gcaactcgac tacacgaagc tggaatcgct1321 agtaatcgcg aatcagaatg tcgcggtgaa tacgttcccg ggtcttgtac acaccgcccg1381 tcacaccatg ggagttggta atgcccgaag tcagtgaccc aaccgcaagg agggagctgc1441 cgaaggcagg actgataact ggggtgaagt cgtaacaagg gtacg(consensus 16S rRNA sequence for Roseburia hominis strain 433)SEQ ID NO: 3AAGAGTTTGGGHCAGGCTCAGGATGAACGCTGGCGGCGTGCTTAACACATGCAAGTCGAACGAAGCACTTTAATTGATTTCTTCGGAATGAAGTTTTTGTGACTGAGTGGCGGACGGGTGAGTAACGCGTGGGTAACCTGCCTCATACAGGGGGATAACAGTTGGAAACGACTGCTAATACCGCATAAGCGCACAGGATTGCATGATCCAGTGTGAAAAACTCCGGTGGTATGAGATGGACCCGCGTCTGATTAGCCAGTTGGCGGGGTAACGGCCCACCAAAGCGACGATCAGTAGCCGACCTGAGAGGGTGACCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGGGAAACCCTGATGCAGCGACGCCGCGTGAGCGAAGAAGTATTTCGGTATGTAAAGCTCTATCAGCAGGGAAGAAGAATGACGGTACCTGACTAAGAAGCACCGGCTAAATACGTGCCAGCAGCCGCGGTAATACGTATGGTGCAAGCGTTATCCGGATTTACTGGGTGTAAAGGGAGCGCAGGCGGTACGGCAAGTCTGATGTGAAATCCCGGGGCTCAACCCCGGTACTGCATTGGAAACTGTCGGACTAGAGTGTCGGAGGGGTAAGTGGAATTCCTAGTGTAGCGGTGAAATGCGTAGATATTAGGAGGAACACCAGTGGCGAAGGCGGCTTACTGGACGATTACTGACGCTGAGGCTCGAAAGCGTGGGGAGCAAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGATGAATACTAGGTGTCGGGGAGCATTGCTCTTCGGTGCCGCAGCAAACGCAATAAGTATNCCACCTGGGGAGTACGTTCGCAAGAATGAAACTCAAAGGAATTGACGGGGACCCGCACAAGCGGTGGAGCNTGTGGTTTAATTCGAAGCAACGCGAAGAACCTTACCAAGTCTTGACATCCCACTGACAGAGTATGTAATGTACTTTCTCTTCGGAGCAGTGGTGACAGGTGGTGCATGGTTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCCTATTCTTAGTAGCCAGCGGTTTGGCCGGGCACTCTAGGGAGACTGCCAGGGATAACCTGGAGGAAGGTGGGGATGACGTCAAATCATCATGCCCCTTATGACTTGGGCTACACACGTGCTACAATGGCGTAAACAAAGGGAAGCAATCCCGCGAGGGGGAGCAAATCTCAAAAATAACGTCTCAGTTCGGACTGTAGTCTGCAACTCGACTACACGAAGCTGGAATCGCTAGTAATCGCGAATCAGAATGTCGCGGTGAATACGTTCCCGGGTCTTGTACACACCGCCCGTCACACCATGGGAGTTGGTAATGCCCGAAGTCAGTGACCCAACCGCAAGGAGGGAGCTGCCGAAGGCAGGACTGATAACTGGGGTGAAGTCTACRSAGGGTAGCCGTRMMC.(Consensus 16S rRNA sequence, Strain A Roseburia intestinalis)SEQ ID NO: 5gctccctcct tgcggttggg tcactgactt cgggcattac caactcccat ggtgtgacgg   60gcggtgtgta caagacccgg gaacgtattc accgcgacat tctgattcgc gattactagc  120gattccagct tcgtgcagtc gagttgcaga ctgcagtccg aactgagacg ttatttttga  180gatttgctcc ccctcgcagg ctcgcttccc tttgtttacg ccattgtagc acgtgtgtag  240cccaagtcat aaggggcatg atgatttgac gtcatcccca ccttcctcca ggttatccct  300ggcagtctcc ctagagtgcc cggcttaccc gctggctact aagaataggg gttgcgctcg  360ttgcgggact taacccaaca tctcacgaca cgagctgacg acaaccatgc accacctgtc  420accgatgctc cgaagagaaa acacattaca tgttctgtca tcgggatgtc aagacttggt  480aaggttcttc gcgttgcttc gaattaaacc acatgctcca ccgcttgtgc gggtccccgt  540caattccttt gagtttcatt cttgcgaacg tactccccag gtggaatact tattgcgttt  600gctgcggcac cgaagagcaa tgctccccga cacctagtat tcatcgttta cggcgtggac  660taccagggta tctaatcctg tttgctcccc acgctttcga gcctcagcgt cagtaatcgt  720ccagtaagcc gccttcgcca ctggtgttcc tcctaatatc tacgcatttc accgctacac  780taggaattcc acttacccct ccgacactct agtccgacag tttccaatgc agtaccgggg  840ttgagccccg ggctttcaca tcagacttgc cgtaccgcct gcgctccctt tacacccagt  900aaatccggat aacgcttgca ccatacgtat taccgcggct gctggcacgt atttagccgg  960tgcttcttag tcaggtaccg tcatttcttc ttccctgnct gatagagctt tacataccga 1020aatacttctt cgctcacgcg gcgtcgctgc atcagggttt cccccattgt gcaatattcc 1080ccactgctgc ctcccgtagg agtttgggcc gtgtctcagt cccaatgtgg ccggtcaccc 1140tctcaggtcg gctactgatc gtcgctttgg taggccgtta ccccaccaac tggctaatca 1200gacgcgggtc catctcatac caccggagtt tttcacacca ggtcatgcga ccctgtgcgc 1260ttatgcggta ttagcagtcg tttccaactg ttatccccct gtatgaggca ggttacccac 1320gcgttactca cccgtccgcc actcagtcac aaaatcttca ttccgaagaa atcaaataaa 1380gtgcttcgtt cgactgca                                               1398(Consensus 16S rRNA sequence, Strain B Roseburia faecis) SEQ ID NO: 6agctccctcc ttgcggttgg gtcactgact tcggacattt ccaactccca tggtgtgacg   60ggcggtgtgt acaagacccg ggaacgtatt caccgcagca ttctgatctg cgattactag  120cgattccagc ttcgtgtagt cgggttgcag actacagtcc gaactgagac gttatttttg  180agatttgctc ggcctcacgg ctttgcttcc ctttgtttac gccattgtag cacgtgtgta  240gcccaagtca taaggggcat gatgatttga cgtcatcccc gccttcctcc aggttatccc  300tggcagtctc cctagagtgc ccggccgaac cgctggctac taaggacagg ggttgcgctc  360gttgcgggac ttaacccaac atctcacgac acgagctgac gacaaccatg caccacctgt  420caccgatgct ccgaagagaa agtacattac atactctgtc atcgggatgt caagacttgg  480taaggttctt cgcgttgctt cgaattaaac cacatgctcc accgcttgtg cgggtccccg  540tcaattcctt tgagtttcat tcttgcgaac gtactcccca ggtggaatac ttattgcgtt  600tgctgcggca ccgaagagca atgctccccg acacctagta ttcatcgttt acggcgtgga  660ctaccagggt atctaatcct gtttgctccc cacgctttcg agcctcagcg tcagttatcg  720tccagtaagc cgccttcgcc actggtgttc ctcctaatat ctacgcattt caccgctaca  780ctaggaattc cacttacccc tccgacactc tagtacgaca gtttccaatg cagtaccggg  840gttgagcccc gggctttcac atcagacttg ccgcaccgcc tgcgctccct ttacacccag  900taaatccgga taacgcttgc accatacgta ttaccgcggc tgctggcacg tatttagccg  960gtgcttctta gtcaggtacc gtcattcttc ttccctgctg atagagcttt acataccgaa 1020atacttcttc gctcacgcgg cgtcgctgca tcagggtttc ccccattgtg caatattccc 1080cactgctgcc tcccgtagga gtttgggccg tgtctcagtc ccaatgtggc cggtcaccct 1140ctcaggtcgg ctactgatcg tcgctttggt aggccgttac cctgccaact ggctaatcag 1200acgcgggtcc atctcatacc accggagttt ttcacaccgg atcatgcgat cctgtgcgct 1260tatgcggtat tagcagtcgt ttccaactgt tatccccctg tatgaggcag gttacccacg 1320cgttactcac ccgtccgcca ctcagtcaca aaatcttcat ttccgaagaa aatcaaatag 1380agtgcttcgt ccgactgcag                                             1400

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1.-24. (canceled)
 25. A composition comprising a bacteria strain of thegenus Roseburia, for use in a method of treating Parkinson's disease,progressive supranuclear palsy, Steele-Richardson-Olszewski syndrome,normal pressure hydrocephalus, vascular parkinsonism, arterioscleroticparkinsonism, drug-induced parkinsonism, Alzheimer's disease, Benson'ssyndrome, Huntington's disease, amyotrophic lateral sclerosis, LouGehrig's disease, motor neurone disease, prion disease, spinocerebellarataxia, spinal muscular atrophy, dementia, Lewy body dementia, vasculardementia, frontotemporal dementia, primary progressive aphasia, mildcognitive impairment, HIV-related cognitive impairment, or corticobasaldegeneration in a subject in need thereof.
 26. The composition of claim25, wherein said composition is for use in a method of treating anearly-onset Parkinson's disease, Alzheimer's disease, Huntington'sdisease, amyotrophic lateral sclerosis, Lou Gehrig's disease, motorneurone disease, prion disease, spinocerebellar ataxia, spinal muscularatrophy, dementia, primary progressive aphasia, mild cognitiveimpairment, HIV-related cognitive impairment, or corticobasaldegeneration.
 27. The composition of claim 25, wherein said compositionis for use in a method of delaying an onset or a progression ofParkinson's disease, Alzheimer's disease, Huntington's disease,amyotrophic lateral sclerosis, Lou Gehrig's disease, motor neuronedisease, prion disease, spinocerebellar ataxia, spinal muscular atrophy,dementia, primary progressive aphasia, mild cognitive impairment,HIV-related cognitive impairment, or corticobasal degeneration.
 28. Thecomposition of claim 25, wherein said bacteria strain is of the speciesRoseburia hominis.
 29. The composition of claim 28, wherein saidbacteria strain comprises a 16S rRNA gene sequence that has at least 96%sequence identity to the polynucleotide sequence of SEQ ID NO:3. 30.(canceled)
 31. (canceled)
 32. The composition of claim 25, wherein saidcomposition is for use in a method of treating Parkinson's disease. 33.(canceled)
 34. (canceled)
 35. The composition of claim 25, wherein saidbacteria strain is lyophilized.
 36. The composition of claim 25, whereinsaid composition is for oral administration.
 37. (canceled)
 38. Acomposition comprising a bacteria strain of the genus Roseburia, for usein a method of treating a stroke in a subject in need thereof, whereinsaid stroke is selected from the group consisting of cerebral ischemia,focal cerebral ischemia, an ischemic stroke, and a hemorrhagic stroke.39. The composition of claim 38, wherein said bacteria strain is of thespecies Roseburia hominis.
 40. The composition of claim 39, wherein saidbacteria strain comprises a 16S rRNA gene sequence that has at least 96%sequence identity to the polynucleotide sequence of SEQ ID NO:3. 41.(canceled)
 42. The composition of claim 38, wherein said bacteria strainis lyophilized.
 43. The composition of claim 38, wherein saidcomposition is for oral administration.
 44. (canceled)